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WO2025217275A2 - Immune cell targeted compositions and related methods - Google Patents

Immune cell targeted compositions and related methods

Info

Publication number
WO2025217275A2
WO2025217275A2 PCT/US2025/023849 US2025023849W WO2025217275A2 WO 2025217275 A2 WO2025217275 A2 WO 2025217275A2 US 2025023849 W US2025023849 W US 2025023849W WO 2025217275 A2 WO2025217275 A2 WO 2025217275A2
Authority
WO
WIPO (PCT)
Prior art keywords
nucleotides
seq
nucleotide sequence
igll5
antisense strand
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/US2025/023849
Other languages
French (fr)
Inventor
Simon Felix BRUNNER
Zeynep Kalender ATAK
Gerda KILDISIUTE
Matthew Daniel YOUNG
Timothy Michael BUTLER
Heather Elizabeth MACHADO
Jacob Rosenblum RUBENS
Geoffrey A. Von Maltzahn
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Flagship Pioneering Innovations VII Inc
Original Assignee
Flagship Pioneering Innovations VII Inc
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Filing date
Publication date
Application filed by Flagship Pioneering Innovations VII Inc filed Critical Flagship Pioneering Innovations VII Inc
Publication of WO2025217275A2 publication Critical patent/WO2025217275A2/en
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]

Definitions

  • RNA agents e.g., double stranded RNAi agents comprising a sense strand and an antisense strand
  • agents e.g., RNA agents (e.g., double stranded RNAi agents comprising a sense strand and an antisense strand) targeting one or more genes expressed by one or more subsets of immune cells, e.g., T cells, B-cells.
  • the disclosure further relates to pharmaceutical compositions comprising the same; and methods of utilizing the same, including, e.g., methods of inhibiting expression of one or more target gene expressed by one or more subsets of immune cells e.g., T cells, B-cells, etc.), and methods of treating diseases associated with expression of one or more of said genes (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer) comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.).
  • diseases associated with expression of one or more of said genes e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer
  • cancer comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.).
  • Somatic mutations are nucleotide alterations in a genomic DNA sequence (e.g., of a gene) that are acquired during the lifetime of a subject. Unlike germline mutations, somatic mutations are not present in the germline DNA of an individual and are therefore not inherited from a parent like germline polymorphisms. Somatic mutations may occur spontaneously due to e.g., the infidelity of DNA replication occurring at each cell division creating substitutions, deletions, or additions of nucleotides into the DNA of a cell. A somatic mutation may also be caused by environmental factors such as e.g., ultraviolet radiation, chemical exposure, or virial infections.
  • environmental factors such as e.g., ultraviolet radiation, chemical exposure, or virial infections.
  • agents e.g., RNA agents (e.g., RNAi agents, dsRNA agents, ssRNA agents, siRNA agents, antisense oligonucleotides, etc.) targeting one or more genes expressed by one or more subsets of immune cells, e.g., T cells, B-cells, tumor infiltrating lymphocytes (TILs) (e.g., tumor infiltrating T-cells); and methods of manufacturing and pharmaceutical compositions comprising the same.
  • RNA agents e.g., RNAi agents, dsRNA agents, ssRNA agents, siRNA agents, antisense oligonucleotides, etc.
  • TILs tumor infiltrating lymphocytes
  • RNA agents e.g., RNAi agents, dsRNA agents, etc.
  • methods of inhibiting or decreasing gene expression e.g., mRNA expression
  • methods of treating diseases associated with the one or more genes including, e.g., proinflammatory diseases (e.g., autoimmune diseases); and cancer; and methods of treating diseases associated with expression of one or more of said genes (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer) comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.).
  • proinflammatory diseases e.g., autoimmune diseases
  • cancer comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.).
  • double stranded ribonucleic acid (dsRNA) agents for inhibiting expression of BTG1 e.g., human BTG1 (hBTGl)
  • BTG1 e.g., human BTG1 (hBTGl)
  • the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
  • dsRNA agents for inhibiting expression of BTG1 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding BTG1 (e.g., (hBTGl).
  • the region of complementarity is fully complementary to the portion of the mRNA sequence encoding BTG1 (e.g., (hBTG1 ). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
  • dsRNA agents for inhibiting expression of BTG1 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding BTG1 (e.g., (hBTGl), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
  • dsRNA double stranded ribonucleic agents for inhibiting expression of CDKN1B (e.g., human CDKN1B (hCDKNIB)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
  • CDKN1B e.g., human CDKN1B (hCDKNIB)
  • hCDKNIB human CDKN1B
  • the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
  • arc dsRNA agents for inhibiting expression of CDKN1B comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding CDKN1B (e.g., (hCDKNIB).
  • the region of complementarity is fully complementary to the portion of the mRNA sequence encoding CDKN1B (e.g., (hCDKNIB). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
  • dsRNA agents for inhibiting expression of CDKN1B comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding CDKN1B (e.g., (hCDKNIB), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
  • double stranded ribonucleic (dsRNA) agents for inhibiting expression of DUSP2 e.g., human DUSP2 (hDUSP2)
  • DUSP2 e.g., human DUSP2 (hDUSP2)
  • the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e. ., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
  • dsRNA agents for inhibiting expression of DUSP2 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding DUSP2 (e.g., (hDUSP2), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
  • DUSP2 e.g., (hDUSP2)
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
  • dsRNA agents for inhibiting expression of DUSP2 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding DUSP2 (e.g., (hDUSP2).
  • the region of complementarity is fully complementary to the portion of the mRNA sequence encoding DUSP2 (e.g., (hDUSP2)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
  • dsRNA agents for inhibiting expression of DUSP2 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding DUSP2 (e.g., (hDUSP2), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
  • DUSP2 e.g., (hDUSP2)
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
  • dsRNA double stranded ribonucleic
  • IGLL5 e.g., human IGLL5 (hIGLL5)
  • antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
  • dsRNA agents for inhibiting expression of IGLL5 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding IGLL5 (e.g., (hIGLL5).
  • the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 (e.g., (hIGLL5)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
  • dsRNA agents for inhibiting expression of IGLL5 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
  • dsRNA double stranded ribonucleic agents for inhibiting expression of IGLL5 (e.g., human IGLL5 (hIGLL5)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
  • IGLL5 e.g., human IGLL5 (hIGLL5)
  • the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
  • arc dsRNA agents for inhibiting expression of IGLL5 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding IGLL5 e.g., (hIGLL5).
  • the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 e.g., (hIGLL5)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
  • dsRNA agents for inhibiting expression of IGLL5 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA sequence encoding IGLL5 (e.g., (hIGLL5), wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 14.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region sequence of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
  • double stranded ribonucleic (dsRNA) agents for inhibiting expression of KLHL6 e.g., human KLHL6 (hKLHL6)
  • KLHL6 e.g., human KLHL6 (hKLHL6)
  • the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
  • dsRNA agents for inhibiting expression of KLHL6 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding KLHL6 (e.g., (hKLHL6).
  • the region of complementarity is fully complementary to the portion of the mRNA sequence encoding KLHL6 (e.g., (hKLHL6)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
  • dsRNA agents for inhibiting expression of KLHL6 comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding KLHL6 (e.g., (hKLHL6), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
  • KLHL6 e.g., (hKLHL6)
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.1n some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 (e.g., (hIGLL5)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides.
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
  • the send strand comprises at least one modified nucleotide and/or the antisense strand comprises at least one modified nucleotide.
  • the sense strand comprises at least one modified nucleotide and/or the antisense strand comprises at least one modified nucleotide. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and/or antisense strand are modified. In some embodiments, substantially all (or all) of the nucleotides in the sense strand and/or antisense strand are modified. In some embodiments, at least one of the modified nucleotides comprises a modified sugar (e.g., ribose moiety).
  • a modified sugar e.g., ribose moiety
  • At least one of the modified nucleotides comprises a modified nucleobase.
  • the sense strand comprises at least one modified internucleoside linkage and/or the antisense strand comprises at least one modified intemucleoside linkage.
  • each of the antisense strand and the sense strand are not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides in length.
  • the antisense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-30 20- 30, 21-30, 22-30, 23-30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21 , 19-22, 19-23, 19-24, or 19-25 nucleotides; and/or the sense strand comprises from about 15-30, 16-30, 17-30,
  • the antisense strand comprises or consists of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides; and/or the antisense strand comprises or consist of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides. In some embodiments, antisense strand comprises from about 19-23 nucleotides; and/or the sense strand comprises from about 19-23 nucleotides. In some embodiments, antisense strand comprises or consists of about 23 nucleotides; and/or the sense strand comprises or consists of about 21 nucleotides.
  • the sense strand and/or the antisense strand comprises a 3' and/or 5' overhang of 1, 2, or 3 nucleotides.
  • the antisense strand comprises a 3' overhang of 1, 2, or 3 nucleotides (e.g., 2 nucleotides).
  • the double stranded region is from about 19-30, 19-29, 19-28,
  • the double stranded region is from about 19-23 or 19-21 nucleotide pairs in length. In some embodiments, the double stranded region is about 21 nucleotide pairs in length.
  • the sense strand and the antisense strand are part of a single nucleic acid molecule (e.g., wherein a hairpin loop is between the sense strand and the antisense strand of the single nucleic acid molecule).
  • the sense strand and the antisense strand are separate nucleic acid molecules (i.e., connected only through the double stranded region).
  • conjugates comprising a dsRNA agent described herein and a heterologous moiety.
  • the heterologous moiety is a peptide, protein, carbohydrate, lipid, polymer, or small molecule.
  • the heterologous moiety is attached to the dsRNA agent via a linker.
  • the heterologous moiety attached to the 3' end of the sense and/or antisense strand and/or the 5' end of the sense and/or antisense strand, and/or at an internal site of the sense and/or antisense strand.
  • vectors e.g., a viral vector, a non-viral vector
  • RNA agent e.g., dsRNA agent
  • carriers comprising a dsRNA agent described herein, a conjugate described herein, or a vector described herein.
  • the carrier comprises a nanoparticle, a polymer, a lipid-based delivery system, as dendrimer, a cationic delivery system, or a hydrogel.
  • the lipid-based delivery system is a lipid nanoparticle (LNP), liposome, lipoplex, nanoliposome, an exosome, or a micelle.
  • LNP lipid nanoparticle
  • cells comprising a dsRNA agent described herein, a conjugate described herein, a vector described herein, or a carrier described herein.
  • compositions comprising a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a cell described herein, and a pharmaceutically acceptable excipient.
  • kits comprising a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein.
  • diagnostic kits comprising one or more reagent for use in a method of detecting the presence or absence of one or more somatic BTG1 (e.g., hBTGl), CDKN1B, DUSP2, IGLL5, or KLHL6 mutation in a sample from a subject.
  • somatic BTG1 e.g., hBTGl
  • CDKN1B CDKN1B
  • DUSP2 DUSP2
  • IGLL5 IGLL5
  • a gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 associated disease in a subject
  • the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and (c) administering to the subject a therapeutic agent if the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA,
  • the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
  • the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
  • the gene associated disease is a BTG1 associated disease, a CDKN1B associated disease, DUSP2 associated disease, a ILL5 associated disease, or a KLHL6 associated disease.
  • the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein
  • the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein
  • the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein
  • the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or
  • the therapeutic agent comprises an anti-inflammatory agent (e.g., an anti-inflammatory agent described herein).
  • an anti-inflammatory agent e.g., an anti-inflammatory agent described herein.
  • the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
  • methods of treating, ameliorating, or preventing a proinflammatory disease in a subject comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and (c) administering to the subject a therapeutic agent if the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
  • somatic gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • the gene is BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6.
  • the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
  • the therapeutic agent is an anti-inflammatory agent as described herein.
  • the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
  • kits for diagnosing a gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in a subject comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the presence of the one or more somatic mutation (e.g., a somatic mutation set forth in Table 2) indicates that the subject has a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease.
  • a gene e.g., BTG1, CDKN1B, DUSP2, IGLL
  • the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
  • the one or more somatic mutation is one or more somatic mutation set forth herein.
  • the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
  • the gene is associated disease is a BTG1 associated disease, a CD KN IB associated disease, DUSP2 associated disease, a IGLL5 associated disease, or a KLHL6 associated disease.
  • the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein
  • the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein
  • the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein
  • the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or
  • the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory (e.g., autoimmune) disease.
  • the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease.
  • (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
  • the method further comprises administering to the subject a therapeutic agent if a somatic mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
  • the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
  • kits for selecting a subject for administration of a therapeutic agent comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the subject is selected for administration of the therapeutic agent if the one or more somatic mutation (e.g., a somatic mutation set forth in Table 2) is present.
  • somatic gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mutation
  • (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
  • the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
  • the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
  • the method further comprises administering to the subject the therapeutic agent if the subject is selected.
  • the therapeutic agent is an antiinflammatory agent as described herein.
  • the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
  • in vitro methods of screening a sample from a subject for one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation e.g., a somatic mutation set forth in Table 2
  • the method comprising (a) isolating and purifying DNA, RNA, or protein from a sample obtained from the subject; and (b) detecting the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
  • (a) comprises isolating and purifying DNA, or having DNA isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
  • the one or more somatic gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
  • the sample is a blood, tissue, or cell sample.
  • the sample is biopsy.
  • the sample comprises a cell (e.g., a T- ccll) or a population of cells (e.g., a population of T-cclls).
  • At least one of the one or more somatic mutation is a loss of function mutation, a gain of function mutation, a missense mutation, synonymous, and/or non- synonymous.
  • the one or more somatic mutation is a loss of function mutation.
  • the one or more somatic mutation truncates the protein encoded by the gene.
  • the one or more somatic mutation is one or more somatic mutation set forth herein.
  • the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
  • a gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • the method comprising delivering into the cell the a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell.
  • a gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject.
  • the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
  • a gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 associated disease in a subject
  • the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby treat, ameliorate, or prevent the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in the subject.
  • the gene associated disease is a BTG1 associated disease, a CDKN1B associated disease, a DUSP2 associated disease, an IGLL5 associated disease, or a KLHL6 associated disease.
  • the gene e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 associated disease associated disease is a proinflammatory disease (e.g., an autoimmune disease).
  • the gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 associated disease associated disease is an autoimmune disease.
  • the gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 associated disease is treated, ameliorated, or prevented through the reduction or inhibition expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • a proinflammatory e.g., autoimmune
  • a pharmaceutical composition described herein comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby treat, ameliorate, or prevent the proinflammatory (e.g., autoimmune) disease in the subject.
  • the proinflammatory (e.g., autoimmune) disease is treated, ameliorated, or prevented through a reduction or inhibition of expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
  • a gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • the subject is a human.
  • the inventors have, inter alia, discovered somatic mutations in select genes expressed by subsets of immune cells (e.g., T cells, B-cells, TILs (e.g., tumor infiltrating T-cells), including BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6, particularly within the context of certain diseases, including, e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer.
  • immune cells e.g., T cells, B-cells, TILs (e.g., tumor infiltrating T-cells), including BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6, particularly within the context of certain diseases, including, e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer.
  • a step of detecting (or having detected) one or more somatic mutation in a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6) expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.) is useful in methods of treating e.g., proinflammatory diseases (e.g., autoimmune diseases); and agents capable of inhibiting expression of one or more of these genes (BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) (e.g., RNA agents described herein) are useful in e.g., methods of treating e.g., proinflammatory diseases (e.g., autoimmune diseases).
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6 expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.
  • proinflammatory diseases e.g
  • the current disclosure provides, inter alia, methods of treating diseases associated with expression of one or more of said genes (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer) comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cclls, etc.); agents (e.g., RNA agents (e.g., RNAi agents, dsRNAi agents comprising a sense strand and an antisense strand, ssRNA agents)) capable of inhibiting gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6) expression (e.g., in a cell, e.g., in a cell in a subject) and their use in, inter alia, pharmaceutical compositions, and methods of treating diseases (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer).
  • proinflammatory diseases e.g., autoimmune diseases
  • cancer
  • any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated.
  • administering refers to the physical introduction of an agent (e.g., an RNA agent (e.g., described herein)) e.g., a therapeutic agent (or a precursor of the therapeutic agent that is metabolized or altered within the body of the subject to produce the therapeutic agent in vivo) to a subject, using any of the various methods and delivery systems known to those skilled in the art.
  • Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.
  • agents include, but are not limited polynucleotides (e.g., DNA, RNA (e.g., RNA)), proteins, peptides, small molecules, carbohydrates, lipids, synthetic polymers (e.g., polymers of PEG).
  • polynucleotides e.g., DNA, RNA (e.g., RNA)
  • proteins e.g., proteins, peptides, small molecules, carbohydrates, lipids
  • synthetic polymers e.g., polymers of PEG
  • the term “antisense strand” refers to an RNA molecule (e.g., in some embodiments, part of an RNAi agent (e.g., described herein), part of a dsRNA agent (e.g., described herein)) that comprises a region of complementarity comprising a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to a target nucleic acid sequence (e.g., a target mRNA (e.g., a BTG1 mRNA), a portion of a target mRNA (e.g., a BTG1 mRNA)).
  • a target nucleic acid sequence e.g., a target mRNA (e.g., a BTG1 mRNA), a portion of a target mRNA (e.g., a BTG1 mRNA)
  • the antisense strand is part of a dsRNA molecule that further comprises a sense strand.
  • the antisense strand or antisense oligonucleotide is a ssRNA agent.
  • the term “bicyclic sugar” refers to a modified sugar (e.g., ribose) moiety comprising two rings, wherein the second ring is formed via a bridge connecting two of the atoms in the first ring thereby forming a bicyclic structure.
  • the first ring of the bicyclic sugar moiety is a furanosyl moiety.
  • the furanosyl sugar moiety is a ribosyl moiety.
  • BNA bicyclic nucleoside
  • the term “blunt end” refers to a dsRNA molecule that does not contain any unpaired nucleotides at the end (e.g., 3' terminus, 5' terminus) of the dsRNA molecule (z.e., no nucleotide overhang(s)).
  • the dsRNA molecule can have, for example, a blunt end at the 3' end, 5' end, or both the 3' and 5' end of the molecule.
  • suitable in vivo or vitro conditions e.g., under certain standard conditions, under mammalian (e.g., human) physiological conditions.
  • Complementary sequences include, e.g., Watson-Crick base pairs.
  • complementary nucleobase pairs include adenine (A) and thymine (T); adenine (A) and uracil (U); and cytosine (C) and guanine (G).
  • Complementary nucleobase pairs include natural and modified nucleotides, and nucleotide mimics, at least to the extent that the above hybridization requirements are fulfilled. As such, determinations of complementarity (as described herein) are independent of nucleotide chemical modifications (e.g., as described herein). For example, (C) and 5-methyl cytosine (mC) are both complementary to (G).
  • conjugation refers to chemical conjugation of an RNA agent (e.g., a nucleic acid molecule) with another agent (e.g., carbohydrate, small molecule, polypeptide, polynucleotide, lipid, synthetic polymer (e.g., polymers of polyethylene glycol (PEG)), etc.).
  • the other agent can be directly connected to the RNA agent (e.g., nucleic acid molecule) or indirectly connected through a linker, e.g., as described herein.
  • Chemical conjugation methods are well known in the art, as are commercially available conjugation reagents and kits, with detailed instructions for their use readily available from the commercial suppliers.
  • the term “differing by no more than X nucleotides” and the like (e.g., differing by not more than X nucleotides” in reference to a nucleotide sequence means that the nucleotide sequence comprises no more than X (wherein X is a specified number e.g., 3, 2, 1, 0)) nucleotide variations (as defined herein) relative to a reference sequence.
  • the phrase “wherein the nucleotide sequence of the antisense strand differs by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: X” means that the nucleotide sequence comprises no more than 3 nucleotide variations relative to the nucleotide sequence set forth in the cited SEQ ID NO: X.
  • disease refers to any abnormal condition that impairs physiological function.
  • the term is used broadly to encompass any disorder, illness, abnormality, pathology, sickness, condition, or syndrome in which physiological function is impaired, irrespective of the nature of the etiology.
  • the term disease includes infection (e.g., a viral, bacterial, fungal, protozoal infection).
  • double stranded RNA agent or “dsRNA agent” refers to a complex of two RNA molecules comprising a double stranded region comprising two anti-parallel and at least partially (e.g., substantially, fully) complementary nucleic acid sequences that form the double stranded region.
  • the dsRNA agent comprises a sense strand and an antisense strand.
  • BTG Anti-Proliferation Factor 1 refers to the BTG/transducer of Erb family (Tob) member that functions, e.g., in the regulation of cell cycle progression, cell proliferation, and apoptosis.
  • BTG1 includes human BTG1 (hBTGl).
  • the mRNA sequence of reference hBTGl gene is set forth in SEQ ID NO: 1 (NCBI Ref.: NM_001731.3).
  • the amino acid sequence of a reference hBTGl protein is set forth in SEQ ID NO: 3 (NCBI Ref.: NP_001722.1).
  • BTGI includes naturally occurring variants of BTG1.
  • BTG1 gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
  • cyclin-dependent kinase inhibitor IB or “CDKN1B” refers to the cyclin-dependent kinase inhibitor that belongs to the Cip/Kip family of cyclin dependent kinase inhibitor proteins.
  • CDKN1B includes human CDKN1B (hCDKNIB).
  • the mRNA sequence of reference hCDKNIB gene is set forth in SEQ ID NO: 4 (NCBI Ref.: NM_004064.5).
  • the amino acid sequence of a reference hCDKNIB protein is set forth in SEQ ID NO: 6 (NCBI Ref.: NP_004055.1).
  • CDKN1B includes naturally occurring variants of CDKN1B.
  • CDKN1B gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
  • DUSP2 dual specificity phosphatase 2
  • DUSP2 refers to the dual specificity protein phosphatase that functions, e.g., in the inactivation of ERK1/2 and p38 MAPK.
  • the term DUSP2 includes human DUSP2 (hDUSP2).
  • the mRNA sequence of reference hDUSP2 gene is set forth in SEQ ID NO: 7 (NCBI Ref.: NM_004418.4).
  • the amino acid sequence of a reference hDUSP2 protein is set forth in SEQ ID NO: 9 (NCBI Ref.: NP_004409.1).
  • DUSP2 includes naturally occurring variants of DUSP2.
  • DUSP2 gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
  • immunoglobulin lambda like polypeptide 5 or “IGLL5” refers to the gene (and encoded protein) located within the immunoglobulin lambda locus, wherein the first exon of the IGLL5 gene is unrelated to immunoglobulin variable genes; while the second and third exons are the immunoglobulin lambda joining 1 and the immunoglobulin lambda constant 1 gene segments.
  • the term IGLL5 includes human IGLL5 (hIGLL5).
  • the mRNA sequence of reference hIGLL5 gene is set forth in SEQ ID NO: 10 (NCBI Ref.: NM_001178126.2).
  • the amino acid sequence of a reference hIGLL5 protein (isoform 1) is set forth in SEQ ID NO: 12 (NCBI Ref.: NP_001171597.1).
  • the mRNA sequence of reference hIGLL5 gene (isoform 2) is set forth in SEQ ID NO: 4 (NCBI Ref.: NM_001256296.2).
  • the amino acid sequence of a reference hIGLL5 protein (isoform 2) is set forth in SEQ ID NO: 15 (NCBI Ref.: NP_001243225.1).
  • the term IGLL5 includes naturally occurring variants of IGLL5.
  • IGLL5 gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
  • non-human primate e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)
  • GenBank GenBank
  • UniProt UniProt
  • OMIM Macaca fascicularis
  • the mRNA sequence of reference hKLHL6 gene is set forth in SEQ ID NO: 16 (NCBI Ref.: NM_ 130446.4).
  • the amino acid sequence of a reference hKLHL6 protein is set forth in SEQ ID NO: 18 (NCBI Ref.: NP_569713.2).
  • KLHL6 includes naturally occurring variants of KLHL6.
  • KLHL6 gene and mRNA sequences of e.g., human, mouse, rat, nonhuman primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
  • DNA and “polydeoxyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple deoxyribonucleotides that are polymerized via phosphodiester bonds.
  • Deoxyribonucleotides are nucleotides in which the sugar is deoxyribose.
  • the term “fully complementary” means that in a hybridized pair of a first nucleic acid molecule and a second nucleic acid molecule, 100% (all), of the bases in a contiguous sequence of the first nucleic acid molecule will hybridize with the same number of bases in a contiguous sequence of the second nucleic acid molecule.
  • the contiguous sequence may comprise all or a part of the first and/or second nucleic acid molecule.
  • nucleic acid molecule comprising a “heterologous moiety” means a nucleic acid molecule that is joined to a moiety (e.g., carbohydrate, small molecule, polypeptide, polynucleotide, lipid, synthetic polymer (e.g., polymers of PEG), etc.) that is not joined to the nucleic acid molecule in nature.
  • isolated with reference to a polypeptide, protein, or polynucleotide refers to a polypeptide, protein, or polynucleotide that is substantially free of other cellular components with which it is associated in the natural state.
  • nucleotide variation refers to a nucleic acid molecule that comprises at least one substitution, addition, or deletion of one or more nucleotide compared to a reference nucleic acid molecule.
  • variant or variant with reference to a peptide or protein refers to a peptide or protein that comprises at least one substitution, addition, or deletion of an amino acid residue compared to a reference peptide or protein.
  • modified RNA agent refers to any RNA agent (or any component thereof (e.g., any nucleic acid molecule thereof)) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) described herein that comprises one or more modified nucleotide (as defined herein).
  • modified nucleotide refers to a nucleotide comprising a chemical modification, e.g., a modified sugar moiety, a modified nucleobase, and/or a modified internucleoside linkage, or any combination thereof.
  • exemplary modifications are provided herein, see, e.g., ⁇ 4.3, 4.3.1.
  • RNA agent e.g., RNAi agent
  • component thereof e.g., described herein, e.g., a sense strand, an antisense strand, a dsRNA agent
  • nucleotide overhang refers to at least one unpaired nucleotide that extends from the double stranded region of a nucleic acid molecule (e.g., a dsRNA molecule (e.g., a dsRNA molecule described herein)). For example, when a 3'-end of one strand of a dsRNA extends beyond the 5'-end of the other strand, or vice versa, there is a nucleotide overhang.
  • non-complementary nucleotide mismatch refers to a nucleotide within a region of complementarity (as described herein) that is not complementary to the corresponding nucleotide in the target nucleic acid molecule.
  • the term “obtaining a sample” refers to the acquisition of a sample.
  • the term includes the direct acquisition from a subject and the indirect acquisition through one or more third parties wherein one of the third parties directly acquired the sample from the subject.
  • operably connected refers to the linkage of two moieties in a functional relationship.
  • a polypeptide is operably connected to another polypeptide when they are linked (either directly or indirectly via a peptide linker) in frame such that both polypeptides are functional (e.g., a fusion protein described herein).
  • a transcription regulatory polynucleotide e.g., a promoter, enhancer, or other expression control element is operably linked to a polynucleotide that encodes a protein if it affects the transcription of the polynucleotide that encodes the protein.
  • the term “operably connected” can also refer to the conjugation of a moiety to e.g., a polynucleotide or polypeptide (e.g., the conjugation of a PEG polymer to a protein).
  • partially complementary means that in a hybridized pair of a first nucleic acid molecule and a second nucleic acid molecule, at least 70%, but not all, of the bases in a contiguous sequence of the first nucleic acid molecule will hybridize with the same number of bases in a contiguous sequence of the second nucleic acid molecule.
  • the contiguous sequence may comprise all or a part of a first or second nucleic acid molecule.
  • Gapped BEAST can be utilized as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, which is incorporated herein by reference in its entirety.
  • PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.).
  • the default parameters of the respective programs e.g., of XBLAST and NBLAST
  • NCBI National Center for Biotechnology Information
  • Another specific, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17, which is incorporated herein by reference in its entirety.
  • ALIGN program version 2.0 which is part of the GCG sequence alignment software package.
  • a PAM 120 weight residue table a gap length penalty of 12
  • a gap penalty of 4 a gap penalty of 4.
  • the percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.
  • composition means a composition that is suitable for administration to an animal, e.g., a human subject, and comprises a therapeutic agent and a pharmaceutically acceptable carrier or diluent.
  • the term “pharmaceutically acceptable carrier or diluent” means a substance intended for use in contact with the tissues of human beings and/or non-human animals, and without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable therapeutic benefit/risk ratio.
  • nucleic acid molecule refers to a polymer of DNA or RNA.
  • the nucleic acid molecule can be single- stranded or double-stranded; contain natural, non-natural, or modified nucleotides; and contain a natural, non-natural, or modified intemucleoside linkage, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule.
  • Nucleic acid molecules include, but are not limited to, all nucleic acid molecules which are obtained by any means available in the art, including, without limitation, recombinant means, e.g., the cloning of nucleic acid molecules from a recombinant library or a cell genome, using ordinary cloning technology and polymerase chain reaction, and the like, and by synthetic means.
  • recombinant means e.g., the cloning of nucleic acid molecules from a recombinant library or a cell genome
  • synthetic means e.g., the cloning of nucleic acid molecules from a recombinant library or a cell genome, using ordinary cloning technology and polymerase chain reaction, and the like, and by synthetic means.
  • recombinant means e.g., the cloning of nucleic acid molecules from a recombinant library or a cell genome, using ordinary cloning technology and polymerase chain reaction, and the like, and
  • any of the RNA polynucleotides encoded by a DNA identified by a particular sequence identification number may also comprise the corresponding RNA (e.g., mRNA) sequence encoded by the DNA, where each thymidine (T) of the DNA sequence is substituted with uracil (U).
  • RNA e.g., mRNA
  • T thymidine
  • U uracil
  • the term “plurality” means 2 or more (e.g., 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 9 or more, or 10 or more).
  • the terms “protein” and “polypeptide” refers to a polymer of at least 2 (e.g., at least 5) amino acids linked by a peptide bond.
  • the term “polypeptide” does not denote a specific length of the polymer chain of amino acids. It is common in the art to refer to shorter polymers of amino acids e.g., approximately 2-50 amino acids) as peptides; and to refer to longer polymers of amino acids (e.g., approximately over 50 amino acids) as polypeptides.
  • the terms “peptide” and “polypeptide” and “protein” are used interchangeably herein.
  • the protein is folded into its three-dimensional structure.
  • proteins folded into their three-dimensional structure are also provided herein as well as polypeptides in the primary structure. Proteins can include more than one polypeptide (e.g., customarily referred to as the quaternary structure).
  • prognostication refers to the process of estimating/predicting the likely course and/or outcome of a disease in a subject, including, e.g., the chance that a subject has of recovering from the disease.
  • region of complementarity refers to a portion of a first nucleic acid molecule comprising a nucleotide sequence that is at least partially complementary to the nucleotide sequence of at least a portion of a second nucleic acid molecule.
  • RNA and “polyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple ribonucleotides that are polymerized via phosphodiester bonds. Ribonucleotides are nucleotides in which the sugar’ is ribose. RNA may contain modified nucleotides; and contain natural, non-natural, or modified internucleoside linkages, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule.
  • RNAi agent refers to an RNA agent that contains one or more RNA molecules which can mediate the targeted cleavage of an RNA molecule (e.g., an mRNA molecule) via an RNA-induced silencing complex (RISC) pathway.
  • RISC RNA-induced silencing complex
  • the RNAi agent is thereby capable of e.g., modulating, e.g., inhibiting, the expression of a target gene (e.g., BTG1) in a cell, e.g., a cell within a subject, such as a mammalian subject.
  • a target gene e.g., BTG1
  • the RNAi agent is a dsRNA agent comprising a sense strand and an antisense strand that form a double stranded region.
  • the sense strand and the antisense strand each independently comprise or consist of from about 19-23 nucleotides.
  • sample encompasses a variety of biological specimens obtained from a subject.
  • sample types include, e.g., blood, red blood cells, and other liquid samples of biological origin (including, but not limited to, whole-blood, red blood cells e.g., isolated from whole blood), peripheral blood mononuclear' cells (PBMCs), serum, plasma, urine, saliva, amniotic fluid, stool, synovial fluid, etc.), nasopharyngeal swabs, solid tissue samples such as biopsies (or cells derived therefrom and the progeny thereof), tissue cultures (or cells derived therefrom and the progeny thereof), and cell cultures (or cells derived therefrom and the progeny thereof).
  • PBMCs peripheral blood mononuclear' cells
  • the term also includes samples that have been manipulated in any way after their procurement from a subject, such as by centrifugation, filtration, washing, precipitation, dialysis, chromatography, lysis, treatment with reagents, enriched for certain cell populations, refrigeration, freezing, staining, etc.
  • the term “sense strand” refers to an RNA molecule (e.g., part of an RNAi agent (e.g., described herein), part of a dsRNA agent (e.g., described herein)) that comprises a region that is at least partially (e.g., substantially, fully) complementary to a region of the antisense strand (as defined herein).
  • the sense strand is often referred to as such with reference to the orientation of the sequence of the sense strand being the same with respect to a target RNA (e.g., mRNA sequence).
  • somatic mutation refers to one or more nucleotide alteration (e.g., variation, modification (e.g., variation)) in a genomic DNA sequence (e.g., of a gene) that is acquired during the lifetime of a subject. It is understood in the art that somatic mutations are not present in the germline DNA of an individual and are therefore not inherited from a parent like germline polymorphisms. Somatic mutations may occur spontaneously due to e.g., the infidelity of DNA replication occurring at each cell division creating substitutions, deletions, or additions of nucleotides into the DNA of a cell. A somatic mutation may also be caused by environmental factors such as e.g., ultraviolet radiation, chemical exposure, or virial infections.
  • environmental factors such as e.g., ultraviolet radiation, chemical exposure, or virial infections.
  • the term “subject” includes any animal, such as a human or other animal.
  • the subject is a vertebrate animal (e.g., mammal, bird, fish, reptile, or amphibian).
  • the subject is a human.
  • the method subject is a non-human mammal.
  • the subject is a non-human mammal is such as a non-human primate (e.g., monkeys, apes), ungulate (e.g., cattle, buffalo, sheep, goat, pig, camel, llama, alpaca, deer, horses, donkeys), carnivore (e.g., dog, cat), rodent (e.g., rat, mouse), or lagomorph (e.g., rabbit).
  • a non-human primate e.g., monkeys, apes
  • ungulate e.g., cattle, buffalo, sheep, goat, pig, camel, llama, alpaca, deer, horses, donkeys
  • carnivore e.g., dog, cat
  • rodent e.g., rat, mouse
  • lagomorph e.g., rabbit
  • the subject is a bird, such as a member of the avian taxa Galliformes (e.g., chickens, turkeys, pheasants, quail), Anseriformes (e.g., ducks, geese), Paleaognathae (e.g., ostriches, emus), Columbiformes (e.g., pigeons, doves), or Psittaciformes (e.g., parrots).
  • avian taxa Galliformes e.g., chickens, turkeys, pheasants, quail
  • Anseriformes e.g., ducks, geese
  • Paleaognathae e.g., ostriches, emus
  • Columbiformes e.g., pigeons, doves
  • Psittaciformes e.g., par
  • substantially complementary means that in a hybridized pair of a first nucleic acid molecule and a second nucleic acid molecule, at least 85%, but not all, of the bases in a contiguous sequence of the first nucleic acid molecule will hybridize with the same number of bases in a contiguous sequence of the second nucleic acid molecule.
  • the contiguous sequence may comprise all or a pail of a first or second nucleic acid molecule.
  • the term “substantially all” means at least 95%, 96%, 97%, 98% or 99%, e.g., of the subject of said sentence, but excluding 100% (i.e., all).
  • target nucleic acid sequence refers to a contiguous portion of the nucleotide sequence of a nucleic acid sequence (e.g., an mRNA molecule formed during the transcription of a target gene (e.g., BTG1)).
  • the target nucleic acid sequence is an mRNA molecule formed during the transcription of a target gene (e.g., BTG1)).
  • the target nucleic acid molecule comprises an mRNA that is a product of RNA processing of a primary transcription product.
  • the target portion of the sequence (e.g., mRNA) will be at least long enough to serve as a substrate for RNAi-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a BTG1 gene.
  • the target sequence is within the protein coding region of BTG1.
  • the term “therapeutically effective amount” of a therapeutic agent refers to any amount of the therapeutic agent that, when used alone or in combination with another therapeutic agent, improves a disease condition, e.g., protects a subject against the onset of a disease; improves a symptom of disease, e.g., decreases severity of disease symptoms, decreases frequency or duration of disease symptoms, increases disease symptom- free periods; prevents or reduces impairment or disability due to the disease; or promotes disease (or infection) regression.
  • a therapeutic agent to improve a disease condition can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the RNA agent in in vitro assays.
  • the terms “treat,” treating,” “treatment,” and the like refer to reducing or ameliorating a disease and/or symptom(s) associated therewith or obtaining a desired pharmacologic and/or physiologic effect. It will be appreciated that, although not precluded, treating a disease does not require that the disease, or symptom(s) associated therewith be completely eliminated. In some embodiments, the effect is therapeutic, i.e., without limitation, the effect partially or completely reduces, diminishes, abrogates, abates, alleviates, decreases the intensity of, or cures a disease and/or adverse symptom attributable to the disease.
  • the effect is preventative, i.e., the effect protects or prevents an occurrence or reoccurrence of a disease.
  • the presently disclosed methods comprise administering a therapeutically effective amount of a compositions as described herein.
  • agents useful in, inter alia, inhibiting expression of one or more target gene (e.g., expressed by one or more subsets of immune cells, e.g., T cells) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of mRNA encoded by the target gene).
  • target gene e.g., expressed by one or more subsets of immune cells, e.g., T cells
  • Target genes include, BTG1 (e.g., human BTG1 (hBTGl)); CDKN1B (e.g., human CDKN1B (hCDKNIB)); DUSP2 (e.g., human DUSP2 (hDUSP2)); IGLL5 (e.g., human IGLL5 (hIGLL5)); or KLHL6 (e.g., human KLHL6 (hKLHL6)).
  • BTG1 e.g., human BTG1 (hBTGl)
  • CDKN1B e.g., human CDKN1B (hCDKNIB)
  • DUSP2 e.g., human DUSP2 (hDUSP2)
  • IGLL5 e.g., human IGLL5 (hIGLL5)
  • KLHL6 e.g., human KLHL6 (hKLHL6)
  • RNA agents useful in, inter alia, inhibiting expression of: BTG1 (BTG1) (e.g., human BTG1 (hBTGl)) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of BTG1 (e.g., hBTGl) mRNA); CDKN1B (e.g., hCDKNIB) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of CDKN
  • BTG1 BTG1
  • hBTGl human BTG1
  • CDKN1B e.g., hCDKNIB
  • BTG1 is a member of the BTG/ Tob family, generally regarded as an anti-proliferative gene family.
  • the BTG/Tob family consists of six protein members; BTG1, BTG2/Tis21/PC3, BTG3/ANA, BTG4/PC3B, TOB1/TOB, and TOB2.
  • BTG1 functions in a wide range of cellular activities including, for example, proliferation, apoptosis, and cell growth.
  • BTG1 expression is generally highest in the G0/G1 phases of the cell cycle and downregulated as cells progress through Gl.
  • BTG1 is expressed both in the nucleus and cytoplasm, and function influenced at least in part by the intracellular localization.
  • BTG1 In the nuclease BTG1 is known to interact with several nuclear receptors and can function as a coactivator of cell differentiation. See, e.g., Kim, Sang Hyeon et al. “Emerging role of anti-proliferative protein BTG1 and BTG2.” BMB reports vol. 55,8 (2022): 380-388. doi:10.5483/BMBRep.2022.55.8.092.
  • the mRNA sequence of a reference hBTGl gene is set forth in SEQ ID NO: 1.
  • the reverse complement sequence of the hBTGl mRNA is set forth in SEQ ID NO: 2.
  • the amino acid sequence of the hBTGl protein encoded by the hBTGl reference gene is set forth in SEQ ID NO: 3.
  • CDKN1B is a cyclin-dependent kinase inhibitor that belongs to the Cip/Kip family of cyclin dependent kinase inhibitor proteins.
  • CDKN1B functions to, e.g., bind to and prevent the activation of cyclin E-CDK2 or cyclin D-CDK4 complexes, and thus control the cell cycle progression at the Gl phase.
  • CDKN1B degradation which is triggered by, e.g., its CDK dependent phosphorylation and subsequent ubiquitination by SCF complexes (Skp, Cullin, F-box containing complex), is generally needed for the cellular transition from quiescence to a proliferative state.
  • the mRNA sequence of a reference hCDKNIB gene is set forth in SEQ ID NO: 4.
  • the reverse complement sequence of the hCDKNIB mRNA is set forth in SEQ ID NO: 5.
  • the amino acid sequence of the hCDKNIB protein encoded by the hCDKNIB reference gene is set forth in SEQ ID NO: 6.
  • DUSP2 is a member of the dual specificity protein phosphatase subfamily. These phosphatases inactivate their target kinases by dephosphorylating both phosphoserine/threonine and pho sphotyro sine residues. Members of the DUSP family exhibit distinct substrate specificities for various MAP kinases, different tissue distribution, different subcellular localization, and different modes of inducibility of their expression by extracellular stimuli. DUSP2, for example, inactivates ERK1/2 and p38 MAPK. DUSP2 is expressed in, e.g., hematopoietic tissues, and is localized in the nucleus.
  • the mRNA sequence of a reference hDUSP2 gene is set forth in SEQ ID NO: 7.
  • the reverse complement sequence of the hDUSP2 mRNA is set forth in SEQ ID NO: 8.
  • the amino acid sequence of the hDUSP2 protein encoded by the hDUSP2 reference gene is set forth in SEQ ID NO: 9.
  • the IGLL5 gene is located within the immunoglobulin lambda locus but it does not require somatic rearrangement for expression.
  • the first exon of the IGLL5 gene is unrelated to immunoglobulin variable genes; while the second and third exons are the immunoglobulin lambda joining 1 and the immunoglobulin lambda constant 1 gene segments. It is known that alternative splicing results in multiple transcript variants.
  • the mRNA sequence of a reference hIGLL5 gene (isoform 1) is set forth in SEQ ID NO: 10.
  • the reverse complement sequence of the hIGLL5 mRNA is set forth in SEQ ID NO: 11.
  • the amino acid sequence of the hIGLL5 protein encoded by the hIGLL5 (isoform 1) reference gene is set forth in SEQ ID NO: 12.
  • the mRNA sequence of a reference hIGLL5 gene is set forth in SEQ ID NO: 13.
  • the reverse complement sequence of the hIGLLS mRNA is set forth in SEQ ID NO: 14.
  • the amino acid sequence of the hIGLL5 protein encoded by the hIGLL5 (isoform 2) reference gene is set forth in SEQ ID NO: 15.
  • KLHL6 is a member of the kelch-like (KLHL) family of proteins. Proteins of the KLHL family generally contain a BTB/POZ domain, a BACK domain, and five to six Kelch motifs.
  • the KLHL6 protein contains an N-terminal broad-complex, tramtrack and brie a brae (BTB) domain that facilitates protein binding and dimerization, a BTB domain, C-terminal kelch (BACK) domain, and six C-terminal kelch repeat domains.
  • BTB broad-complex, tramtrack and brie
  • BTB brae
  • BACK C-terminal kelch
  • KLHL6 functions, e.g., in B-lymphocyte antigen receptor signaling and germinal-center B-cell maturation.
  • KLHL6 is expressed by lymphoid tissue, including, e.g., by germinal center B-cells.
  • the mRNA sequence of a reference hKLHL6 gene is set forth in SEQ ID NO: 16.
  • the reverse complement sequence of the hKLHL6 mRNA is set forth in SEQ ID NO: 17.
  • the amino acid sequence of the hKLHL6 protein encoded by the hKLHL6 reference gene is set forth in SEQ ID NO: 18.
  • the RNA agent comprises one or more RNA molecule.
  • the RNA agent is an RNAi agent.
  • the RNAi agent is a dsRNA agent.
  • the RNA agent is an siRNA agent.
  • the RNA agent comprises a dsRNA agent.
  • the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand.
  • the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand that form a double stranded region.
  • the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand that hybridize to form a double stranded region.
  • the sense strand and the antisense strand are part of a single nucleic acid molecule (e.g., a single nucleic acid molecule comprising a hairpin loop).
  • the sense strand and the antisense strand are separate nucleic acid molecules.
  • the RNA agent comprises an antisense strand.
  • the RNA agent comprises a sense strand.
  • the RNA agent comprises one or more single stranded RNA (ssRNA) molecule.
  • the RNA agent consists of an antisense strand.
  • the RNA agent is an antisense oligonucleotide agent.
  • the RNA agent is an RNAi agent.
  • the RNAi agent comprises an antisense strand and a sense strand.
  • the RNAi agent is a double stranded RNA (dsRNA) agent comprising a sense strand and an antisense strand.
  • the RNAi agent is an siRNA agent.
  • Antisense strands comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least substantially complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is fully complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, 1GLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, 1GLL5, or KLHL6 mRNA
  • a portion of a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 70% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 75% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 80% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 85% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 90% complementary to the nucleotide sequence of the target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 95% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least 95%, 96%, 97%, 98%, 99%, or 100% (e.g., in some embodiments, preferably at least 95%, more preferably at least 98%) complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity comprises or consists of one or more non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity comprises or consists of no more than 5 (e.g., 4, 3, 2, 1, or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 5 (e.g., 4, 3, 2, 1 , or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 3 (e.
  • nucleotide sequence of the region of complementarity comprises or consists of no more than 2 (e.g., 1 or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 1 (e.g., 0) non-complementary nucleotide mismatch relative to the nucleotide sequence of the target nucleic acid molecule.
  • the nucleotide sequence of the region of complementarity comprises 0 non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule.
  • the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity.
  • the region of complementarity comprises at least one but not more than 3 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non- complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity.
  • the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity.
  • the last 3 e.g., 2 or 1
  • the region of complementarity comprises at least one but not more than 3 non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity.
  • a target nucleic acid molecule e.g., a target mRNA (e.g., a BTGl , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA))).
  • a target mRNA e.g., a BTGl , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the region of complementarity comprises from about 15-30 nucleotides, e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30,
  • the region of complementarity comprises from about 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-
  • the region of complementarity comprises from about 19-21 (e.g., 19-20) nucleotides. In some embodiments, the region of complementarity comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity comprises about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity comprises about 19 nucleotides. In some embodiments, the region of complementarity comprises about 20 nucleotides.
  • the region of complementarity comprises about 21 nucleotides. In some embodiments, the region of complementarity comprises about 22 nucleotides. In some embodiments, the region of complementarity comprises about 23 nucleotides. In some embodiments, the region of complementarity consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23,
  • the region of complementarity consists of about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity consists of about 19 nucleotides. In some embodiments, the region of complementarity consists of about 20 nucleotides. In some embodiments, the region of complementarity consists of about 21 nucleotides. In some embodiments, the region of complementarity consists of about 22 nucleotides. In some embodiments, the region of complementarity consists of about 23 nucleotides. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
  • the target nucleic acid molecule is part (e.g., a contiguous portion) of a larger nucleic acid molecule.
  • the target nucleic acid molecule is a portion (e.g., a contiguous portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the target nucleic acid molecule is a contiguous nucleotide sequence of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) of sufficient length to allow it to be a substrate for cleavage directed by an RNAi agent e.g., an RNAi agent described herein, e.g., a dsRNA agent (e.g., described herein)) (i.e., cleavage through a RISC pathway).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the target nucleic acid molecule is a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA). In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) formed in the expression of a target gene (e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene) (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 gene).
  • a target gene e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene
  • a target gene e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a BTG1 (e.g., hBTGl) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a BTG1 (e.g., hBTGl) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 1 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a BTG1 (e.g., hBTGl) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 3 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a CDKNIB (e.g., hCDKNIB) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a CDKNIB (e.g., hCDKNIB) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 4 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a CDKN1B e.g., hCDKNIB) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 6 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a DUSP2 (e.g., hDUSP2) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a DUSP2 (e.g., hDUSP2) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 7 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a DUSP2 (e.g., hDUSP2) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 9 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a IGLL5 (e.g., hIGLL5) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a IGLL5 (e.g., hIGLL5) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 10 (or a valiant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 12 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 13 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein.
  • IGLL5 e.g., hIGLL5
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 15 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a KLHL6 (e.g., hKLHL6) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a KLHL6 (e.g., hKLHL6) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 16 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a KLHL6 (e.g., hKLHL6) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 18 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises from about 19-30 nucleotides, e.g., 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20- 29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26,
  • the target nucleic acid molecule comprises from about 19-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 19-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 19, 18, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 19 nucleotides.
  • the target nucleic acid molecule comprises about 20 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 21 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 23 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 19, 18, 20,
  • the target nucleic acid molecule consists of about 19 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 20 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 21 nucleotides. Tn some embodiments, the target nucleic acid molecule consists of about 23 nucleotides.
  • the antisense strand comprises from about 15-30 nucleotides (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17,
  • the antisense strand comprises from about 18-25 nucleotides (e.g., 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20- 24,20-23, 20-22, 20-21 , 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides).
  • the antisense strand comprises from about 19-25 nucleotide (e.g., 19-20, 19-21, 19-22, 19-23, 19-24, 19-25, 20-21, 20-22, 20-23, 20-24, 20-25, 21-22, 21-23,
  • the antisense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-3020-30, 21-30, 22-30, 23- 30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides. In some embodiments, the antisense strand comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense strand comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
  • the antisense strand comprises about 21 nucleotides. In some embodiments, the antisense strand comprises about 23 nucleotides. In some embodiments, the antisense strand consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense strand consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense strand consists of about 21 nucleotides. In some embodiments, the antisense strand consists of about 23 nucleotides.
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises at least 30 (e.g., at least 31, 32,
  • the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of a portion of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,
  • nucleotide 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 , 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 19 contiguous nucleotides.
  • the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20 contiguous nucleotides.
  • the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 21 contiguous nucleotides.
  • the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 22 contiguous nucleotides.
  • the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 23 contiguous nucleotides.
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 19 contiguous nucleotides.
  • the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20 contiguous nucleotides.
  • the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 21 contiguous nucleotides.
  • the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 22 contiguous nucleotides.
  • the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 23 contiguous nucleotides.
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 20 e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense strand comprises 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 19 contiguous nucleotides.
  • the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20 contiguous nucleotides.
  • the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 21 contiguous nucleotides.
  • the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 22 contiguous nucleotides.
  • the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 23 contiguous nucleotides.
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 15, 16, 17, 18,
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,
  • the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27,
  • the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
  • the antisense strand consists of
  • the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28,
  • the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11 , 14, or 17. In some embodiments, the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 ⁇ e.g., 0, 1, 2, 3, 4, or 5) ⁇ e.g., by no more than 3 ⁇ e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 19 contiguous nucleotides.
  • the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 ⁇ e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 ⁇ e.g., 0, 1, 2, 3, 4, or 5) ⁇ e.g., by no more than 3 ⁇ e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 20 contiguous nucleotides.
  • the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 ⁇ e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 ⁇ e.g., 0, 1, 2, 3, 4, or 5) ⁇ e.g., by no more than 3 ⁇ e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 21 contiguous nucleotides.
  • the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 ⁇ e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 ⁇ e.g., 0, 1, 2, 3, 4, or 5) ⁇ e.g., by no more than 3 ⁇ e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 22 contiguous nucleotides.
  • the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 23 contiguous nucleotides.
  • sense strands comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • pairs of sense and antisense strands can hybridize to form a double stranded region (e.g., under conditions in which the pairs will be used).
  • the nucleotide sequence of the region of complementarity is at least substantially complementary to the nucleotide sequence of at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity is fully complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity may be at least 70% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity may be at least 75% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity may be at least 80% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity may be at least 85% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity may be at least 90% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity may be at least 95% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity is at least 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity comprises one or more non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 5 (e.g., 4, 3, 2, 1, or 0) non- complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity comprises no more than 3 (e.g., 2, 1, or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 2 e.g., 1 or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand.
  • the nucleotide sequence of the region of complementarity comprises no more than 1 (e.g., 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises 0 non- complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand.
  • the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand, wherein the one or more non- complementary nucleotide mismatch is within the last 5 (e.g., 4, 3, 2, or 1 ) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity.
  • the last 5 e.g., 4, 3, 2, or 1
  • the region of complementarity comprises at least one but not more than 3 (e.g., 1, 2, or 3) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand, wherein the one or more non-complementary nucleotide mismatch is within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity.
  • 3 e.g., 1, 2, or 3
  • the region of complementarity comprises from about 15-30 nucleotides, e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30,
  • the region of complementarity comprises from about 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20- 24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides.
  • the region of complementarity comprises from about 19-21 (e.g., 19-20) nucleotides. In some embodiments, the region of complementarity comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity comprises about 19, 20, or 21 nucleotides. In some embodiments, the region of complementarity comprises about 19 nucleotides. In some embodiments, the region of complementarity comprises about 20 nucleotides. In some embodiments, the region of complementarity comprises about 21 nucleotides. In some embodiments, the region of complementarity consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
  • the region of complementarity consists of about 19, 20, or 21 nucleotides. In some embodiments, the region of complementarity consists of about 19 nucleotides. In some embodiments, the region of complementarity consists of about 20 nucleotides. In some embodiments, the region of complementarity consists of about 21 nucleotides.
  • the sense strand comprises from about 15-30 nucleotides (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21 , 15-20, 15-19, 15-18, 15-17, 18- 30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28,
  • 15-30 nucleotides e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21 , 15-20, 15-19, 15-18, 15-17, 18- 30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28,
  • the sense strand comprises from about 18-25 nucleotides (e.g.,
  • the sense strand comprises from about 19-25 nucleotide (e.g.,
  • the sense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-30 20-30, 21-30, 22-30, 23-30, 24- 30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides. [00216] In some embodiments, the sense strand comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense strand comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
  • the sense strand comprises about 19, 20, 21, 22, 23 nucleotides. In some embodiments, the sense strand comprises about 19, 20, 21 nucleotides. In some embodiments, the sense strand comprises about 20 nucleotides. In some embodiments, the sense strand comprises about 21 nucleotides. In some embodiments, the sense strand comprises about 21 nucleotides. In some embodiments, the sense strand comprises about 23 nucleotides.
  • the sense strand consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense strand consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense strand consists of about 19, 20, 21, 22, 23 nucleotides. In some embodiments, the sense strand consists of about 19, 20, 21 nucleotides. In some embodiments, the sense strand consists of about 20 nucleotides. In some embodiments, the sense strand consists of about 21 nucleotides. In some embodiments, the sense strand consists of about 21 nucleotides. In some embodiments, the sense strand consists of about 23 nucleotides.
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the sense strand comprises at least 30 e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a B TGI, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 19 contiguous nucleotides.
  • the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 20 contiguous nucleotides.
  • the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 21 contiguous nucleotides.
  • the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 22 contiguous nucleotides.
  • the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand comprises 23 contiguous nucleotides.
  • the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7.
  • the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16.
  • the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 19 contiguous nucleotides.
  • the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 20 contiguous nucleotides.
  • the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 21 contiguous nucleotides.
  • the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 22 contiguous nucleotides.
  • the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the sense strand consists of 23 contiguous nucleotides.
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the sense strand comprises at least 25 (e.g., at least 26, 27 , 28,
  • the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29,
  • the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35 contiguous nucleotides.
  • the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 ⁇ e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 19 contiguous nucleotides. [00245] In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20 contiguous nucleotides.
  • the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1 , 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
  • the sense strand comprises 21 contiguous nucleotides. [00247] In some embodiments, the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
  • the sense strand comprises 22 contiguous nucleotides.
  • the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
  • the sense strand comprises 23 contiguous nucleotides. [00249] In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1 , 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,
  • the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,
  • the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 19 contiguous nucleotides. [00252] In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20 contiguous nucleotides.
  • the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
  • the sense strand consists of 21 contiguous nucleotides. [00254] In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
  • the sense strand consists of 22 contiguous nucleotides. [00255] In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
  • the sense strand consists of 23 contiguous nucleotides.
  • the RNA agent (e.g., RNAi agent) comprises a dsRNA agent comprising an antisense strand (e.g., described herein, e.g., described in ⁇ 4.2.1.1) and a sense strand (e.g., described herein, e.g., described in ⁇ 4.2.1.2) that hybridize to form a double stranded region (e.g., under conditions in which the dsRNA will be used (e.g., under physiological (e.g., mammalian, e.g., human) conditions within a cell)).
  • a dsRNA agent comprising an antisense strand (e.g., described herein, e.g., described in ⁇ 4.2.1.1) and a sense strand (e.g., described herein, e.g., described in ⁇ 4.2.1.2) that hybridize to form a double stranded region (e.g., under conditions in which the dsRNA will
  • antisense strands comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)); and the sense strands comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of at least a portion of an antisense strand.
  • a target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP
  • the sense strand and the antisense strand can be part of a single larger nucleic acid molecule (connected as a single stranded nucleic acid molecule) or separate nucleic acid molecules (only connected through the double stranded region).
  • the sense strand and the antisense strand are separate nucleic acid molecules.
  • sense strand and the antisense strand are pail of a single larger nucleic acid molecule.
  • the nucleic acid molecule may comprise a hairpin loop between the antisense strand and the sense strand to allow for formation of the double stranded region.
  • the hairpin loop comprises at least 1 (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 23, 25 or more) unpaired nucleotides (non-complementary nucleotide mismatches).
  • the hairpin loop comprises at least one but less than 25, 23, 20, 10, 9, 8, 7, 6, 5, 4, 3, or 2 unpaired nucleotides (non-complementary nucleotide mismatches).
  • the hairpin loop comprises about 25, 23, 20, 9, 8, 7, 6, 5, 4, 3, or 1 unpaired nucleotide (non-complementary nucleotide mismatch). In some embodiments, the hairpin loop consists of about 25, 23, 20, 9, 8, 7, 6, 5, 4, 3, or 1 unpaired nucleotide (non-complementary nucleotide mismatch).
  • the nucleic acid molecule after introduction into a suitable cell (e.g., a mammalian cell, e.g., a human cell), the nucleic acid molecule may be cleaved into a dsRNA molecule wherein the two strands of the dsRNA molecule are no longer part of the same nucleic acid molecule e.g., by a Type III endonuclease (e.g., Dicer) (see, e.g., Sharp et al. (2001) Genes Dev. 15:485, the entire contents of which are incorporated herein by reference for all purposes).
  • a Type III endonuclease e.g., Dicer
  • the double stranded region is about 15-30 base pairs in length (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17,
  • the double stranded region is about 18-25 base pairs in length (e.g., 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20,
  • the double stranded region is about 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15- 20, 15-19, 15-18, 15-17, 15-16, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-20, 19-21, 23-30, 23-29, 23-28, 23-27, 23-26, 23-25, 23-24, 21-30, 21-29, 21-28, 21-27, 21-26,
  • the double stranded region is about 19-21 e.g., 19-20) base pairs in length.
  • the double stranded region is not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length. In some embodiments, the double stranded region is about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length. In some embodiments, the double stranded region is about 19, 20, or 21 base pairs in length. In some embodiments, the double stranded region is about 19 base pairs in length. In some embodiments, the double stranded region is about 20 base pairs in length. In some embodiments, the double stranded region is about 21 base pairs in length. In some embodiments, the double stranded region is about 23 base pairs in length.
  • the double stranded region comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length. In some embodiments, the double stranded region comprises about 19, 20, or 21 base pairs in length. In some embodiments, the double stranded region comprises about 19 base pairs in length. In some embodiments, the double stranded region comprises about 20 base pairs in length. In some embodiments, the double stranded region comprises about 21 base pairs in length. In some embodiments, the double stranded region is about 23 base pairs in length. [00265] In some embodiments, the double stranded region consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length.
  • the double stranded region consists of about 19, 20, or 21 base pairs in length. In some embodiments, the double stranded region consists of about 19 base pairs in length. In some embodiments, the double stranded region consists of about 20 base pairs in length. In some embodiments, the double stranded region consists of about 21 base pairs in length. In some embodiments, the double stranded region is about 23 base pairs in length.
  • the dsRNA agent comprises one or more nucleotide overhang.
  • the nucleotides of a nucleotide overhang can include one or more a modified (e.g., chemically modified) nucleotide (e.g., described herein, e.g., described in ⁇ 4.3, 4.3.1).
  • the nucleotide overhang comprises from about 1-5 nucleotides, e.g., 1-4, 1-3, 1-2, 2-5, 2-4, 2-3, 3-5, 3-4, 4-5 nucleotides. In some embodiments, the nucleotide overhang comprises about 1, 2, 3, 4, or 5 nucleotides. In some embodiments, the nucleotide overhang comprises about 1 nucleotide. In some embodiments, the nucleotide overhang comprises about 2 nucleotides. In some embodiments, the nucleotide overhang consists of about 1, 2, 3, 4, or 5 nucleotides. In some embodiments, the nucleotide overhang consists of about 1 nucleotide. In some embodiments, the nucleotide overhang consists of about 2 nucleotides.
  • the nucleotide overhang(s) can be on the sense strand, the antisense strand, or both the sense strand and the antisense strand.
  • the sense strand comprises a nucleotide overhang.
  • the antisense strand comprises a nucleotide overhang.
  • the sense strand and the antisense strand both comprise a nucleotide overhang.
  • the nucleotide(s) of an overhang can be present on the 5'-end, 3'- end, or both the 5'-end, 3'- end of an antisense and/or sense strand.
  • the sense strand comprises a nucleotide overhang at the 5'-end.
  • the sense strand comprises a nucleotide overhang at the 3'-end.
  • the sense strand comprises a nucleotide overhang at the 5'-end and the 3 '-end.
  • the antisense strand comprises a nucleotide overhang at the 5'-end.
  • the antisense strand comprises a nucleotide overhang at the 3'-cnd. In some embodiments, the antisense strand comprises a nucleotide overhang at the 5'-end and the 3'-end. In some embodiments, the antisense strand comprises a nucleotide overhang at the 3'-end; and the sense strand comprises a nucleotide overhang at the 3'-end. In some embodiments, the antisense strand comprises a nucleotide overhang at the 5'-end; and the sense strand comprises a nucleotide overhang at the 5'-end.
  • the dsRNA agent comprises one or more blunt end.
  • the dsRNA agent comprises a blunt end at the end of the RNA agent comprising the 3 'end of the sense strand and the 5' end of the antisense strand.
  • the dsRNA agent comprises a blunt end at the end of the RNA agent comprising the 5 'end of the sense strand and the 3' end of the antisense strand.
  • both ends of the dsRNA agent are blunt ends.
  • the antisense strand and the sense strand contain the same number of nucleotides. In some embodiments, the antisense strand and the sense strand contain different numbers of nucleotides. In some embodiments, the nucleotide sequence of the sense strand is from about 1-5, 1-3, or 1-2 nucleotides shorter than the nucleotide sequence of the antisense strand. In some embodiments, the nucleotide sequence of the sense strand is about 1, 2, 3, 4, or 5 nucleotides shorter than the nucleotide sequence of the antisense strand.
  • the nucleotide sequence of the sense strand is about 2 nucleotides shorter than the nucleotide sequence of the antisense strand. In some embodiments, the nucleotide sequence of the antisense strand is from about 1-5, 1-3, or 1-2 nucleotides shorter than the nucleotide sequence of the sense strand. In some embodiments, the nucleotide sequence of the antisense strand is about 1, 2, 3, 4, or 5 nucleotides shorter than the nucleotide sequence of the sense strand. In some embodiments, the nucleotide sequence of the antisense strand is about 2 nucleotides shorter than the nucleotide sequence of the sense strand.
  • the sense strand comprises 21 nucleotides. In some embodiments, the antisense strand comprises 23 nucleotides. In some embodiments, the sense strand comprises 21 nucleotides; and the antisense strand comprises 23 nucleotides. In some embodiments, the double stranded region comprises 21 nucleotides. Tn some embodiments, the antisense strand comprises a 2-nuclcotidc overhang at the 3' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand comprises 21 nucleotides; the antisense strand comprises 23 nucleotides; the double stranded region comprises 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand comprises 19 nucleotides.
  • the antisense strand comprises 21 nucleotides.
  • the sense strand comprises 19 nucleotides; and the antisense strand comprises 21 nucleotides.
  • the double stranded region comprises 19 nucleotides.
  • the antisense strand comprises a 2-nucleotide overhang at the 3' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. In some embodiments, the sense strand comprises 19 nucleotides; the antisense strand comprises 21 nucleotides; the double stranded region comprises 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. [00276] In some embodiments, the sense strand comprises 21 nucleotides.
  • the antisense strand comprises 21 nucleotides. In some embodiments, the sense strand comprises 21 nucleotides; and the antisense strand comprises 21 nucleotides. In some embodiments, the double stranded region comprises 19 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3'end. In some embodiments, the sense strand comprises a 2-nucleotide overhang at the 3'end.
  • the sense strand comprises 21 nucleotides; the antisense strand comprises 21 nucleotides; the double stranded region comprises 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 2-nucleotide overhang at the 3' end.
  • the sense strand comprises 20 nucleotides. In some embodiments, the antisense strand comprises 19 nucleotides. In some embodiments, the sense strand comprises 20 nucleotides; and the antisense strand comprises 19 nucleotides. In some embodiments, the double stranded region comprises 20 nucleotides. In some embodiments, the sense strand comprises a 1 -nucleotide overhang at the 5' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand comprises 20 nucleotides; the antisense strand comprises 19 nucleotides; the double stranded region comprises 20 nucleotides; the sense strand comprises a 1 -nucleotide overhang at the 5' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand comprises 21 nucleotides. In some embodiments, the antisense strand comprises 19 nucleotides. In some embodiments, the sense strand comprises 21 nucleotides; and the antisense strand comprises 19 nucleotides. In some embodiments, the double stranded region comprises 19 nucleotides. In some embodiments, the sense strand comprises a 1 -nucleotide overhang at the 3' end. In some embodiments, the sense strand comprises a 1 -nucleotide overhang at the 5' end.
  • the sense strand comprises 21 nucleotides; the antisense strand comprises 19 nucleotides; the double stranded region comprises 19 nucleotides; the sense strand comprises a 1 -nucleotide overhang at the 3' end; and the sense strand comprises a 1 -nucleotide overhang at the 5' end.
  • the sense strand comprises 24 nucleotides. In some embodiments, the antisense strand comprises 23 nucleotides. In some embodiments, the sense strand comprises 24 nucleotides; and the antisense strand comprises 23 nucleotides. In some embodiments, the double stranded region comprises 21 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3 'end. In some embodiments, the sense strand comprises a 3-nucleotide overhang at the 3' end.
  • the sense strand comprises 24 nucleotides; the antisense strand comprises 23 nucleotides; the double stranded region comprises 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 3-nucleotide overhang at the 3' end.
  • the sense strand comprises 19 nucleotides. In some embodiments, the antisense strand comprises 19 nucleotides. In some embodiments, the sense strand comprises 19 nucleotides; and the antisense strand comprises 19 nucleotides. In some embodiments, the double stranded region comprises 19 nucleotides. In some embodiments, the 5' end of the antisense strand (and 3' end of the sense strand) form a blunt end. In some embodiments, the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end.
  • the sense strand comprises 19 nucleotides; the antisense strand comprises 19 nucleotides; the double stranded region comprises 19 nucleotides; the 5' end of the antisense strand (and 3' end of the sense strand) form a blunt end; and the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end
  • the antisense strand and the sense strand are part of the same larger nucleic acid molecule, wherein the nucleic acid molecule comprises 44 nucleotides, the antisense portion comprises 21 nucleotides, the sense strand portion of the nucleic acid molecule comprises 19 nucleotides, the double stranded region comprises 19 nucleotides, the antisense strand comprises a 2-nucleotide overhang at the 3 'end, and the intervening nucleotide sequence between the antisense strand and the sense strand comprises 4 unpaired nucleotides that create a hairpin loop.
  • the sense strand consists of 21 nucleotides. In some embodiments, the antisense strand consists of 23 nucleotides. In some embodiments, the sense strand consists of 21 nucleotides; and the antisense strand consists of 23 nucleotides. In some embodiments, the double stranded region consists of 21 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand consists of 21 nucleotides; the antisense strand consists of 23 nucleotides; the double stranded region consists of 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand consists of 19 nucleotides.
  • the antisense strand consists of 21 nucleotides.
  • the sense strand consists of 19 nucleotides; and the antisense strand consists of 21 nucleotides.
  • the double stranded region consists of 19 nucleotides.
  • the antisense strand comprises a 2-nucleotide overhang at the 3 'end.
  • the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand consists of 19 nucleotides; the antisense strand consists of 21 nucleotides; the double stranded region consists of 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand consists of 21 nucleotides. In some embodiments, the antisense strand consists of 21 nucleotides. In some embodiments, the sense strand consists of 21 nucleotides; and the antisense strand consists of 21 nucleotides. In some embodiments, the double stranded region consists of 19 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3 'end. In some embodiments, the sense strand comprises a 2-nucleotide overhang at the 3'end.
  • the sense strand consists of 21 nucleotides; the antisense strand consists of 21 nucleotides; the double stranded region consists of 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 2-nuclcotidc overhang at the 3' end.
  • the sense strand consists of 20 nucleotides. In some embodiments, the antisense strand consists of 19 nucleotides. In some embodiments, the sense strand consists of 20 nucleotides; and the antisense strand consists of 19 nucleotides. In some embodiments, the double stranded region consists of 20 nucleotides. In some embodiments, the sense strand comprises a 1-nucleotide overhang at the 5' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand consists of 20 nucleotides; the antisense strand consists of 19 nucleotides; the double stranded region consists of 20 nucleotides; the sense strand comprises a 1-nucleotide overhang at the 5' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
  • the sense strand consists of 21 nucleotides. In some embodiments, the antisense strand consists of 19 nucleotides. In some embodiments, the sense strand consists of 21 nucleotides; and the antisense strand consists of 19 nucleotides. In some embodiments, the double stranded region consists of 19 nucleotides. In some embodiments, the sense strand comprises a 1-nucleotide overhang at the 3' end. In some embodiments, the sense strand comprises a 1-nucleotide overhang at the 5' end.
  • the sense strand consists of 21 nucleotides; the antisense strand consists of 19 nucleotides; the double stranded region consists of 19 nucleotides; the sense strand comprises a 1-nucleotide overhang at the 3' end; and the sense strand comprises a 1-nucleotide overhang at the 5' end.
  • the sense strand consists of 24 nucleotides. In some embodiments, the antisense strand consists of 23 nucleotides. In some embodiments, the sense strand consists of 24 nucleotides; and the antisense strand consists of 23 nucleotides. In some embodiments, the double stranded region consists of 21 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3'end. In some embodiments, the sense strand comprises a 3-nucleotide overhang at the 3'end.
  • the sense strand consists of 24 nucleotides; the antisense strand consists of 23 nucleotides; the double stranded region consists of 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 3-nucleotide overhang at the 3' end.
  • the sense strand consists of 19 nucleotides. In some embodiments, the antisense strand consists of 19 nucleotides. In some embodiments, the sense strand consists of 19 nucleotides; and the antisense strand consists of 19 nucleotides. In some embodiments, the double stranded region consists of 19 nucleotides. In some embodiments, the 5' end of the antisense strand (and 3’ end of the sense strand) form a blunt end. In some embodiments, the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end.
  • the sense strand consists of 19 nucleotides; the antisense strand consists of 19 nucleotides; the double stranded region consists of 19 nucleotides; the 5' end of the antisense strand (and 3' end of the sense strand) form a blunt end; and the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end
  • the antisense strand and the sense strand are part of the same larger nucleic acid molecule, wherein the nucleic acid molecule consists of 44 nucleotides, the antisense portion consists of 21 nucleotides, the sense strand portion of the nucleic acid molecule comprises 19 nucleotides, the double stranded region consists of 19 nucleotides, the antisense strand comprises a 2-nucleotide overhang at the 3 'end, and the intervening nucleotide sequence between the antisense strand and the sense strand consists of 4 unpaired nucleotides that create a hairpin loop.
  • the antisense strand is an antisense strand described herein.
  • the sense strand is a sense strand described herein.
  • the antisense strand is an antisense strand described in ⁇ 4.2.1.1.
  • the sense strand is a sense strand described in ⁇ 4.2.1.2.
  • the antisense strand is an antisense strand described in ⁇ 4.2.1.1; and the sense strand is a sense strand described in ⁇ 4.2.1.2.
  • any sense strand described herein e.g., in ⁇ 4.2.1.2
  • any antisense strand in a dsRNA agent described herein e.g., in ⁇ 4.2.1.1
  • the entire contents of in ⁇ 4.2.1.1 and ⁇ 4.2.1.2 are incorporated by reference into the instant section.
  • nucleotide sequence of exemplary unmodified and/or modified dsRNAi agents comprising a sense and antisense strand (e.g., suitable for targeting BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6, suitable for inhibiting BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 expression)) are described in herein.
  • the dsRNA agent (e.g., for inhibiting expression of BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 (e.g., human BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the dsRNA agent e.g., for inhibiting expression of BTG1 (e.g., (hBTGl) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding BTG1 (e.g., (hBTGl), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0,
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1,
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
  • the dsRNA agent (e.g., for inhibiting expression of CDKN1B (e.g., (hCDKNIB) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding CDKN1B (e.g., (hCDKNIB), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucle
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1 , 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 4.
  • 5 e.g., 0, 1 , 2, 3, 4, or 5
  • 3 e.g., 0, 1 , 2, or 3
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
  • no more than 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the dsRNA agent e.g., for inhibiting expression of DUSP2 (e.g., (hDUSP2) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding DUSP2 (e.g., (hDUSP2), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0,
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1,
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
  • the dsRNA agent e.g., for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
  • IGLL5 e.g., (hIGLL5)
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 10.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
  • the dsRNA agent e.g., for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
  • IGLL5 e.g., (hIGLL5)
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 13.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
  • KLHL6 e.g., (hKLHL6)
  • the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 16.
  • the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the dsRNA agent is in a free acid form.
  • the dsRNA agent is in a salt form.
  • the dsRNA agent is in a sodium salt form.
  • sodium ions are present in the composition comprising the dsRNA agent as counterions for substantially all of the phosphodiester or phosphoro thioate groups present in the dsRNA agent.
  • sodium ions are present in the RNA agent as counterions for all of the phosphodiester or phosphorothioate groups present in the dsRNA agent.
  • the RNA agent comprises an antisense oligonucleotide.
  • the antisense oligonucleotide is single stranded.
  • the antisense oligonucleotide is single stranded and is not part of an RNAi agent.
  • Antisense oligonucleotides can mediate the inhibition or alteration of gene expression through, e.g., degradation of a target RNA (e.g., through the recruitment of endogenous enzymes (e.g., RNAses), inhibition of translation of a target RNA (e.g., through steric hinderance), and splicing modulation (e.g., leading to exon skipping, exon inclusion, etc.).
  • endogenous enzymes e.g., RNAses
  • inhibition of translation of a target RNA e.g., through steric hinderance
  • splicing modulation e.g., leading to exon skipping, exon inclusion, etc.
  • antisense oligonucleotides comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least substantially complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is fully complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, 1GLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a portion of a target mRNA e.g., a BTG1, CDKN1B, DUSP2, 1GLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 70% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6
  • the nucleotide sequence of the region of complementarity may be at least 75% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 80% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 85% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 90% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity may be at least 95% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is at least 95%, 96%, 97%, 98%, 99%, or 100% (e.g., in some embodiments, preferably at least 95%, more preferably at least 98%) complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity is 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity comprises or consists of one or more non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the nucleotide sequence of the region of complementarity comprises no more than 5 e.g., 4, 3, 2, 1, or 0) non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 3 (e.g., 2, 1, or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule.
  • the nucleotide sequence of the region of complementarity comprises or consists of no more than 2 (e.g., 1 or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 1 (e.g., 0) non-complementary nucleotide mismatch relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises 0 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule.
  • the region of complementarity comprises one or more (e.g., 2, 3, or more) non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity.
  • the last 5 e.g., 4, 3, 2, or 1
  • the region of complementarity comprises at least one but not more than 3 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity.
  • the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non- complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity.
  • the last 3 e.g., 2 or 1
  • the region of complementarity comprises at least one but not more than 3 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity.
  • a target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA))).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • a portion of a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the region of complementarity comprises from about 15-30 nucleotides, e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30,
  • the region of complementarity comprises from about 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-
  • the region of complementarity comprises from about 19-21 (e.g., 19-20) nucleotides. In some embodiments, the region of complementarity comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity comprises about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity comprises about 19 nucleotides. In some embodiments, the region of complementarity comprises about 20 nucleotides.
  • the region of complementarity comprises about 21 nucleotides. In some embodiments, the region of complementarity comprises about 22 nucleotides. In some embodiments, the region of complementarity comprises about 23 nucleotides. In some embodiments, the region of complementarity consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity consists of about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity consists of about 19 nucleotides. In some embodiments, the region of complementarity consists of about 20 nucleotides. In some embodiments, the region of complementarity consists of about 21 nucleotides.
  • the region of complementarity consists of about 22 nucleotides. In some embodiments, the region of complementarity consists of about 23 nucleotides. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
  • the target nucleic acid molecule is part (e.g., a contiguous portion) of a larger nucleic acid molecule.
  • the target nucleic acid molecule is a portion (e.g., a contiguous portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the target nucleic acid molecule is a contiguous nucleotide sequence of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) of sufficient length to allow it to be a substrate for cleavage directed by the antisense oligonucleotide.
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA
  • the target nucleic acid molecule is a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) formed in the expression of a target gene (e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene) (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 gene).
  • a target gene e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene
  • a target gene e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a BTG1 (e.g., hBTGl) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a BTG1 (e.g., hBTGl) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 1 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a BTG1 (e.g., hBTGl) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 1 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a CDKN1B (e.g., hCDKNIB) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a CDKN 1 B (e.g., hCDKNIB) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 4 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a CDKN1B (e.g., hCDKNIB) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 4 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a DUSP2 (e.g., hDUSP2) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a DUSP2 (e.g., hDUSP2) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 7 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a DUSP2 (e.g., hDUSP2) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 7 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a IGLL5 (e.g., hIGLL5) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a IGLL5 (e.g., hIGLL5) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 10 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 10 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a IGLL5 (e.g., hIGLL5) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a IGLL5 (e.g., hIGLL5) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 13 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 13 (or a variant or fragment thereof).
  • the target nucleic acid molecule is at least a portion (e.g., a portion) of a KLHL6 (e.g., hKLHL6) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a KLHL6 (e.g., hKLHL6) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 16 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a KLHL6 (e.g., hKLHL6) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 16 (or a variant or fragment thereof).
  • the target nucleic acid molecule comprises from about 19-30 nucleotides, e.g., 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20- 29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, 21-22, 22-30, 22-29, 22-28, 22-27, 22-26, 22-25, 22-24, 22-23, 23-30, 23-29, 23-28, 23-27, 23-26, 23-27, 23-26, 23-25, or 23-24 nucleotides.
  • 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22 19-21, 19-20, 20-30, 20- 29, 20-28, 20-27, 20-26, 20-25, 20-24,
  • the target nucleic acid molecule comprises from about 19-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 19-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 19, 18, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the target nucleic acid molecule comprises or consists of about 19 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 20 nucleotides.
  • the target nucleic acid molecule comprises about 21 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 23 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 19, 18, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 19 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 20 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 21 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 23 nucleotides.
  • the antisense oligonucleotide comprises from about 15-50 nucleotides (e.g., 15-49, 15-48, 15-47, 15-46, 15-45, 15-44, 15-43, 15-42, 15-41, 15-40, 15-39, 15-38, 15-37, 15-36, 15-35, 15-34, 15-33, 15-32, 15-31, 15-30, 15-29, 15-28, 15-27, 15- 26, 15- 25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26,
  • 15-50 nucleotides e.g., 15-49, 15-48, 15-47, 15-46, 15-45, 15-44, 15-43, 15-42, 15-41, 15-40, 15-39, 15-38, 15-37, 15-36, 15-35, 15-34, 15-33, 15-32, 15
  • the antisense oligonucleotide comprises from about 15-35 nucleotides (e.g., 15-34, 15-33, 15-32, 15-31, 15-30, 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24,
  • the antisense oligonucleotide comprises from about 15-30 nucleotides (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21 , 21-30, 21 -29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides).
  • 15-30 nucleotides e.g., 15-29, 15-28, 15-27, 15- 26, 15-25,
  • the antisense oligonucleotide comprises from about 18-35 nucleotides (e.g., 18-34, 18-33, 18-32, 18-31, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides). In some embodiments, the antisense oligonucleotide comprises from about 18-30 nucleotides (e.g., 18-34, 18-33, 18-32, 18-31, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22
  • the antisense oligonucleotide comprises from about 18-25 nucleotides (e.g., 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25,
  • the antisense oligonucleotide comprises from about 19-25 nucleotide (e.g., 19-20, 19-21, 19-22, 19-23, 19-24, 19-25, 20-21, 20-22, 20-23, 20-24, 20-25, 21-22, 21-23, 21-24, 21-25, 22-23, 22-24, 22-25, 23-24,
  • the antisense oligonucleotide comprises from about 15-30, 16-30, 17-30, 18-30, 19-3020-30, 21-30, 22-30, 23-30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides.
  • the antisense oligonucleotide comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides. In some embodiments, the antisense oligonucleotide comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides. In some embodiments, the antisense oligonucleotide comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
  • the antisense oligonucleotide comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
  • the antisense oligonucleotide consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides. In some embodiments, the antisense oligonucleotide consists of not more than about 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides.
  • the antisense oligonucleotide consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
  • the antisense oligonucleotide comprises about 18 nucleotides.
  • the antisense oligonucleotide comprises about 19 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 20 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 21 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 22 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 24 nucleotides.
  • the antisense oligonucleotide comprises about 25 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 26 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 27 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 28 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 29 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 31 nucleotides.
  • the antisense oligonucleotide comprises about 32 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 33 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 34 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 35 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 36 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 37 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 38 nucleotides.
  • the antisense oligonucleotide comprises about 39 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 40 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 41 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 42 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 43 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 44 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 45 nucleotides.
  • the antisense oligonucleotide comprises about 46 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 47 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 48 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 49 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 50 nucleotides.
  • the antisense oligonucleotide consists of about 18 nucleotides.
  • the antisense oligonucleotide consists of about 19 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 20 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 21 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 23 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 22 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 23 nucleotides.
  • the antisense oligonucleotide consists of about 24 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 25 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 26 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 27 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 28 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 29 nucleotides.
  • the antisense oligonucleotide consists of about 30 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 31 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 32 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 33 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 34 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 35 nucleotides.
  • the antisense oligonucleotide consists of about 36 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 37 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 38 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 39 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 40 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 41 nucleotides.
  • the antisense oligonucleotide consists of about 42 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 43 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 44 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 45 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 46 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 47 nucleotides.
  • the antisense oligonucleotide consists of about 48 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 49 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 50 nucleotides.
  • the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises at least 30 (e.g., at least 31 , 32, 33, 34, or 35) nucleotides from the nucleotide sequence of a portion of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • no more than 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense oligonucleotide comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • no more than 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 19 contiguous nucleotides.
  • the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides.
  • the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides.
  • the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides.
  • the antisense oligonucleotide comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 23 contiguous nucleotides.
  • the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense oligonucleotide consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 19 contiguous nucleotides.
  • the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides.
  • the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides.
  • the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides.
  • the antisense oligonucleotide consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA.
  • the antisense oligonucleotide consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 23 contiguous nucleotides.
  • the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0,
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8,
  • the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
  • the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,
  • the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25,
  • nucleotide sequence differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides. [00350] In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 21 contiguous nucleotides.
  • the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides.
  • the antisense oligonucleotide comprises 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide comprises 30 contiguous nucleotides differing by no more than I nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 30 contiguous nucleotides.
  • the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • 5 e.g., 0, 1, 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,
  • the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25,
  • nucleotide sequence differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
  • the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides.
  • the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • 5 e.g., 0, 1 , 2, 3, 4, or 5
  • 3 e.g., 0, 1, 2, or 3
  • the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides.
  • the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides.
  • the antisense oligonucleotide consists of 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
  • the antisense oligonucleotide consists of 30 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 30 contiguous nucleotides.
  • the RNA agent e.g., RNAi agent (e.g., siRNA)
  • any component thereof e.g., any nucleic acid molecule thereof
  • the modified agent comprises one or more modified nucleotide(s) (as defined herein).
  • the modified agent may have one or more different (e.g., improved) properties relative to a corresponding unmodified RNA agent.
  • the modified RNA agent may exhibit decreased immunostimulatory activity (e.g., when administered to a subject), increased stability (e.g., in vivo, in a cell, when administered to a subject), and/or increased inhibition of expression of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)), or any combination thereof.
  • a target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)
  • a target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)
  • Nucleotide modifications can include modification to any one of more of the nucleoside and/or the internucleoside linkage. Nucleoside modifications include modification to the sugar (e.g., ribose) moiety and/or the nucleobase.
  • the modified agent (or component thereof) e.g., antisense strand, sense strand, dsRNA agent, etc.
  • the modified agent (or component thereof) comprises one or more nucleotides comprising a modified nucleobase.
  • the modified agent (or component thereof) comprises one or more nucleotides comprising a modified intemucleoside linkage.
  • the modified agent (or component thereof) e.g., antisense strand, sense strand, dsRNA agent, etc.
  • the modified RNA agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, antisense oligonucleotide, etc.) comprises one or more nucleotide comprising a modified nucleoside.
  • nucleoside modifications can include modification of the sugar (e.g., ribose) moiety and/or modification of the nucleobase.
  • the modified RNA agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, antisense oligonucleotide, etc.) comprises one or more nucleotides comprising a modified sugar (e.g., ribose) moiety.
  • a modified sugar e.g., ribose
  • the modified sugar (e.g., ribose) moiety can comprise, for example, a substituent at any one or more position of the sugar (e.g., ribose), including e.g., positions 2', 4', and/or 5'.
  • the modified sugar (e.g., ribose) comprises a substituent at 2' position of the sugar (e.g., ribose).
  • the modified sugar (e.g., ribose) comprises a substituent at 5' position of the sugar (e.g., ribose).
  • the modified sugar (e.g., ribose) comprises a substituent at 5' position of the sugar (e.g., ribose).
  • the RNA agent (or any component thereof) comprises any one or more of the following substituents (e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')): a group for improving the stability of the RNA agent, a group for improving the pharmacokinetic properties of the RNA agent, or a group for improving the pharmacodynamic properties of the RNA agent, an RNA cleaving group, a reporter group, an intercalator, and/or other substituents having similar properties.
  • substituents e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')
  • substituents e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')
  • substituents e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2
  • substituents include, for example, but are not limited to, substitution (e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')) with any one of the following: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl can be substituted or unsubstituted Ci to Cio alkyl or C2 to Cio alkenyl and alkynyl.
  • substitution e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')
  • substitution e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')
  • substitution e.g.,
  • Additional exemplary substitutions include, for example, but are not limited to, substitution with any one of the following: 0[(CH 2 ) n 0]m, CH 3 , O(CH 2 ) U OCH 3 , O(CH 2 ) n NH 2 , O(CH 2 ) n CH 3 , O(CH 2 ) n ONH 2 , and O(CH 2 ) n ON[(CH 2 ) n CH 3 )] 2 , where n and m are from 1 to about 10.
  • the modified sugar comprises any one of the following modifications: 2'-O-methyl (2'-0Me), 2'O-methoxyethyl (2'-0-M0E), 2'deoxy-2'- fluoro (2'-F), 2'-arabino-fluoro (2'-Ara-F), 2'-O-benzyl, 2'-O-methyl-4-pyridine (2-O-methyl-4- pyridine (2'-O-CH2Py(4)).
  • the RNA agent (or any component thereof) comprises any of the following substituents at the 2'-position of the sugar (e.g., ribose): Ci to Cio lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH 3 , OCN, Cl, Br, CN, CF 3 , OCF 3 , SOCH 3 , SO 2 CH 3 , ONO 2 , NO 2 , N 3 , NH 2 , heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, or a substituted silyl.
  • the sugar e.g., ribose
  • the RNA agent (or any component thereof) comprises a 2'-methoxyethoxy (2'-0 — CH2CH2OCH3, also known as 2'-O-(2-mcthoxycthyl) or 2'-M0E) (see, e.g., Martin ct al., Hclv. Chim. Acta, 1995, 78:486-504, the entire contents of which is incorporated by reference herein for all purposes) (i.e., an alkoxyalkoxy group).
  • the RNA agent (or any component thereof) comprises a 2'- dimethylaminooxy ethoxy, i.e., a O(CH2)2ON(CH3)2 group, also known as 2'-DMAOE; a 2'- dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethylaminoethoxyethyl or 2'- DMAEOE), i.e., 2'-O— CH 2 — O— CH 2 — N(CH 3 ) 2 ; a 5'-Me-2'-F nucleotide, a 5'-Me-2'-OMe nucleotide, a 5'-Me-2'-deoxynucleotide, (both R and S isomers in these three families); a 2'- alkoxyalkyl; and 2'-NMA (N-methylacetamide).
  • a 2'- dimethylaminooxy ethoxy i.e.,
  • Exemplary US patents that describe the preparation of such modified sugar structures include, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920; the entire contents of each of the foregoing are hereby incorporated herein by reference for all purposes.
  • the modified sugar (e.g., ribose) moiety comprises a non- bicyclic modified sugar (e.g., ribose) moiety.
  • the modified sugar (e.g., ribose) moiety comprises a furanosyl ring comprising one or more substituent groups none of which bridges two atoms of the furanosyl ring to form a bicyclic structure.
  • one or more non-bridging substituent of a non-bicyclic modified sugar moiety is branched. Such non bridging substituents may be at any position of the furanosyl, including but not limited to substituents at the 2', 4', and/or 5' positions.
  • non-bicyclic modified sugar moiety comprises a substituent group at the 2'-position of the sugar (e.g., ribose).
  • 2'- substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 2'-O-methyl (2'-0Me), 2'0- methoxyethyl (2'-0-M0E), 2 'deoxy-2 '-fluoro (2'-F), 2'-arabino-fluoro (2'-Ara-F), 2'-O-benzyl, 2'- O-methyl-4-pyridine (2-O-methyl-4-pyridine (2'-O-CH2Py(4)), and 2’-O-N-alkyl acetamide (e.g., 2'-O-N-methyl acetamide (“NMA”), 2'-O-N-dimethyl acetamide, 2'-O-N-ethyl acetamide, and 2'- O
  • NMA 2'-O-
  • the 2'-substituent group is a halo, allyl, amino, azido, SH, CN, OCN, CF3, OCF3, O-C1-C10 alkoxy, O-C1-C10 substituted alkoxy, O-C1-C10 alkyl, O-C1-C10 substituted alkyl, S-alkyl, N(R m )-alkyl, O-alkenyl, S-alkenyl, N(R m )-alkenyl, O-alkynyl, S- alkynyl, N(R m )-alkynyl, O-alkylenyl-O- alkyl, alkynyl, alka
  • these 2'-substituent groups can be further substituted with one or more substituent groups independently selected from among: hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO2), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl.
  • a non-bridging 2'-substituent group selected from: F, NH2, N3, OCF3, 0CH3, O(CH2)
  • non-bicyclic modified sugar moiety comprises a substituent group at the 3'-position of the sugar (e.g., ribose).
  • substituent groups suitable for the 3 '-position of modified sugar moieties include but are not limited to alkoxy (e.g., methoxy), alkyl (e.g., methyl, ethyl).
  • non-bicyclic modified sugar moiety comprises a substituent group at the 4'-position of the sugar (e.g., ribose).
  • 4'-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to alkoxy (e.g., methoxy), alkyl, and those described in Manoharan et al., WO 2015/106128.
  • non-bicyclic modified sugar moiety comprises a substituent group at the 5 '-position of the sugar (e.g., ribose).
  • substituent groups suitable for the 5'-position of modified sugar moieties include, but are not limited to, vinyl (e.g., 5'-vinyl), alkoxy (e.g., methoxy (e.g., 5'-methoxy)), and alkyl (e.g., methyl (R or S) (e.g., 5'-methyl (R or S)), ethyl).
  • non-bicyclic modified sugar moieties comprise more than one non-bridging sugar substituent, for example, 2'-F-5'-methyl sugar moieties and the modified sugar moieties and modified nucleosides described in Migawa et al., WO 2008/101157 and Rajeev et al., US2013/0203836, the entire contents of each of which is incorporated herein by reference for all purposes.
  • modified furanosyl sugar moieties and nucleosides incorporating such modified furanosyl sugar moieties are further defined by isomeric configuration.
  • a 2'- deoxyfuranosyl sugar moiety may be in seven isomeric configurations other than the naturally occurring -D- deoxyribosyl configuration.
  • modified sugar moieties are described in, e.g., WO 2019/157531, the entire contents of which are incorporated by reference herein for all purposes.
  • the sugar (e.g., ribose) modification comprises an unlocked nucleotide (UNA).
  • UNA is unlocked acyclic nucleic acid, wherein any of the bonds of the sugar has been removed, forming an unlocked sugar (e.g., ribose) residue.
  • the bonds between Cl'-C4' have been removed (i.e. the covalent carbon-oxygen- carbon bond between the Cl' and C4' carbons).
  • the C2'-C3' bond i.e. the covalent carbon-carbon bond between the C2' and C3' carbons) of the sugar (e.g., ribose) have been removed.
  • the modified sugar’ e.g., ribose
  • the modified sugar comprises a substituent that bridges two atoms of the furanosyl ring to form a second ring, resulting in a bicyclic sugar (e.g., ribose) moiety.
  • the bicyclic sugar e.g., ribose
  • the bicyclic sugar comprises a bridge between the 4’ and the 2’ furanose ring atoms.
  • 4' to 2' bridging sugar substituents include but are not limited to: 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2' (“LNA”), 4'-CH2-S-2’, 4'-(CH2)2-O-2' (“ENA”), 4'-CH(CH3)-O-2' (referred to as “constrained ethyl” or “cEt”), 4'-CH 2 - O-CH 2 -2', 4'-CH 2 -N(R)-2', 4'-CH(CH 2 OCH 3 )-O-2' (“constrained MOE” or “cMOE”) and analogs thereof (see, e.g., Seth et al., U.S.
  • each R, R a , and R b is, independently, H, a protecting group, or C1-C12 alkyl (see, e.g. Imanishi et al., U.S. 7,427,672). The entire contents of all of the foregoing references is incorporated by reference herein for all purposes.
  • the modified sugar comprises a constrained ethyl nucleotide comprising a 4'-CH(CH3) — O-2' bridge.
  • the constrained ethyl nucleotide is in the S conformation (S-cEt).
  • the modified sugar e.g., ribose
  • CRNs are nucleotide analogs with a linker connecting the C2' and C4' carbons of ribose or the C3 and — C5' carbons of ribose. Representative publications that teach the preparation of certain of the above include, but are not limited to, US2013/0190383; and WO2013/036868, the entire contents of each of which are hereby incorporated herein by reference.
  • bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by isomeric configuration.
  • an LNA nucleoside (described herein) may be in the a-L configuration or in the 0-D configuration.
  • general descriptions of bicyclic nucleosides include both isomeric configurations.
  • Any of the foregoing bicyclic nucleosides can be prepared having one or more stereochemical sugar configurations including for example a-L-ribofuranose and -D-ribofuranose (see, e.g., WO 99/14226, the entire contents of which are incorporated herein by reference for all purposes).
  • the modified agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more nucleotides comprising a modified nucleobase.
  • unmodified nucleobases refer to the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Modified nucleobases include other synthetic and natural nucleobases.
  • Modified nucleobases include, but are not limited to, 5-substituted pyrimidines, 6- azapyrimidines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, and N-2, N-6 and 0-6 substituted purines.
  • nucleobases include tricyclic pyrimidines, such as 1,3-diazaphenoxazine- 2-one, l,3-diazaphenothiazine-2-one and 9-(2-aminoethoxy)-l,3-diazaphenoxazine-2-one (G- clamp).
  • Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza- adenine, 7-deazaguanosine, 2- aminopyridine and 2-pyridone.
  • Further nucleobases include those disclosed in Merigan et al., U.S.
  • the modified nucleobase comprises a pseudouridine, 2'thiouridinc (s2U), N6'-mcthyladcnosinc, 5 'mcthylcy tidinc (m 5 C), 5 'fluoro-2 'deoxy uridine, N- ethylpiperidine 7-EAA triazole modified adenine, N-ethylpiperidine 6'triazole modified adenine, 6-phenylpyrrolo-cytosine (PhpC), 2',4'-difluorotoluyl ribonucleoside (rF), or 5 'nitroindole.
  • s2U pseudouridine
  • N6'-mcthyladcnosinc 5 'mcthylcy tidinc
  • m 5 C 5 C
  • 5 'fluoro-2 'deoxy uridine N- ethylpiperidine 7-EAA triazole modified
  • the modified nucleobase comprises a 5-substituted pyrimidine; 6- azapyrimidine; or N-2, N-6 and 0-6 substituted purines (including 2-aminopropyladenine, 5- propynyluracil and 5-propynylcytosine).
  • 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are exemplary base substitutions, even more particularly when combined with 2'-O-methoxyethyl sugar modifications.
  • the modified agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more modified internucleoside linkage.
  • Modified internucleoside linkages compared to naturally occurring phosphate linkages, can be used to alter, typically increase, nuclease resistance of an RNA agent (e.g., described herein).
  • the naturally occurring internucleoside linkage of RNA and DNA is a 3’ to 5' phosphodiester linkage.
  • the modified internucleoside linkage contains a normal 3'-5' linkage.
  • the modified intemucleoside linkage contains a 2'-5' linkage.
  • the modified intemucleoside linkage has an inverted polarity wherein the adjacent pairs of nucleoside units are linked e.g., 3'-5' to 5'-3' or 2'- 5 ' to 5'-2'.
  • the two main classes of modified intcmuclcosidc linking can be defined by the presence or absence of a phosphorous atom.
  • the modified intemucleoside linkage comprises a phosphorous atom.
  • Representative modified phosphorus-containing internucleoside linkages include but are not limited to phosphorothioates (PS (Rp isomer or Sp isomer)) (e.g., 5'phosphorothioate), phosphotriesters, phosphoramidates e.g., 3'-amino phosphoramidate and aminoalky Iphosphoramidates), chiral phosphorothioates, phosphorodithioates (PS2), aminoalkylphosphotriesters, methyl and other alkyl phosphonates (e.g., methylphosphonate (MP), 3'-alkylene phosphonates), methpxypropyl-phosphonates (MOP), 5'-(E)-vinylphosphonates, 5'methyl phosphonates, (S)-5'C-methyl with phosphates, phosphinates, thionophosphoram
  • PS phosphoroth
  • the modified intemucleoside linkage does not contain a phosphorous atom.
  • Modified intemucleoside linkages that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl intemucleoside linkages, mixed heteroatoms and alkyl or cycloalkyl intemucleoside linkages, or one or more short chain heteroatomic or heterocyclic intemucleoside linkages.
  • morpholino linkages formed in part from the sugar portion of a nucleoside
  • siloxane backbones sulfide, sulfoxide and sulfone backbones
  • formacetyl and thioformacetyl backbones methylene formacetyl and thioformacetyl backbones
  • alkene containing backbones sulfamate backbones
  • sulfonate and sulfonamide backbones amide backbones; and others having mixed N, O, S, and CH2 component parts.
  • Methods of preparing polynucleotides comprising modified internucleoside linkages do not contain a phosphorous atom are known in the art. See, e.g., U.S. Pat. Nos.
  • the modified agent comprises one or more RNA mimetic in which both the sugar and the internucleoside linkage of the nucleotide units are replaced with novel groups.
  • the nucleobase units are maintained for hybridization with an appropriate nucleic acid target (e.g., a target mRNA).
  • the RNA mimetic is a peptide nucleic acid (PNA).
  • PNAs the ribose moiety of the RNA nucleotide is replaced with an amide containing moiety (e.g., an aminoethylglycine).
  • the nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide.
  • PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, the entire contents of each of which are hereby incorporated herein by reference. Additional PNA compounds suitable for use in the RNA agents described herein are described in, for example, in Nielsen et al., Science, 1991, 254, 1497-1500, the entire contents of which is incorporated by reference herein for all purposes.
  • RNA agents e.g., described herein
  • modifications to the terminal ends of the RNA agents can also be incorporated to agents described herein.
  • N-(caproyl-4-hydroxyprolinol (Hyp- C6) N-(acetyl-4-hydroxyprolinol (Hyp-NHAc)
  • modifications are known in the art. See, e.g., WO2011/005861 , the entire contents of which is incorporated herein by reference.
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent) are modified.
  • RNA agent or any component e.g., nucleic acid molecule) thereof
  • RNA agent or any component e.g., nucleic acid molecule
  • any component e.g., nucleic acid molecule e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.
  • At least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) arc modified.
  • substantially all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) are modified.
  • all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) are modified.
  • At least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense and/or antisense strand are modified.
  • at least 50% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 55% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 60% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • At least 65% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 70% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 75% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 80% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 85% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 90% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • At least 90% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • at least 95% of the nucleotides of the sense strand and/or antisense strand may be modified.
  • substantially all (or all) of the nucleotides in the sense strand and/or antisense strand are modified.
  • at least one of the modified nucleotides comprises a modified sugar' (e.g., ribose moiety).
  • at least one of the modified nucleotides comprises a modified nucleobase.
  • the sense strand comprises at least one modified internucleoside linkage and/or the antisense strand comprises at least one modified internucleoside linkage.
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent) are unmodified.
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
  • RNA agent 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
  • substantially all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.
  • all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
  • not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.
  • any component e.g., nucleic acid molecule
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified.
  • RNA agent or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified.
  • the RNA agent (e.g., RNAi agent, antisense strand, sense strand, dsRNA agent (e.g., described herein)) comprises one or more non-naturally internucleoside linkage. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the intemucleoside linkages of the RNA agent (e.g., RNAi agent, antisense strand, sense strand, dsRNA agent (e.g., described herein)) are non-naturally occurring.
  • RNA agent e.g., RNAi agent, antisense strand, sense strand, dsRNA agent (e.g., described herein)
  • RNA agent e.g., RNAi agent, antisense strand, sense strand, dsRNA agent (e.g., described herein)
  • dsRNA agent e.g., described herein
  • the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand.
  • a dsRNA agent comprising a sense strand and an antisense strand.
  • at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified.
  • about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified.
  • At least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand are modified. In some embodiments, substantially all of the nucleotides of the sense strand are modified. In some embodiments, all of the nucleotides of the sense strand arc modified.
  • At least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified.
  • at least 50% of the nucleotides of the sense strand may be modified.
  • at least 55% of the nucleotides of the sense strand may be modified.
  • at least 60% of the nucleotides of the sense strand may be modified.
  • at least 65% of the nucleotides of the sense strand may be modified.
  • at least 70% of the nucleotides of the sense strand may be modified.
  • At least 75% of the nucleotides of the sense strand may be modified.
  • at least 80% of the nucleotides of the sense strand may be modified.
  • at least 85% of the nucleotides of the sense strand may be modified.
  • at least 90% of the nucleotides of the sense strand may be modified.
  • at least 90% of the nucleotides of the sense strand may be modified.
  • at least 95% of the nucleotides of the sense strand may be modified.
  • substantially all (or all) of the nucleotides in the sense strand are modified.
  • At least one of the modified nucleotides comprises a modified sugar (e.g., ribose moiety). In some embodiments, at least one of the modified nucleotides comprises a modified nucleobase. In some embodiments, the sense strand comprises at least one modified intemucleoside linkage and/or the antisense strand comprises at least one modified intemucleoside linkage.
  • not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of sense strand are unmodified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand are unmodified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the a sense strand are unmodified.
  • At least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are unmodified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are unmodified.
  • At least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand are unmodified. In some embodiments, substantially all of the nucleotides of the sense strand are unmodified. In some embodiments, all of the nucleotides of the sense strand arc unmodified.
  • not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the sense strand are modified.
  • not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand are modified.
  • not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified.
  • the sense strand comprises one or more non-naturally intemucleoside linkage. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the intemucleoside linkages of sense strand are non-naturally occurring. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the or intemucleoside linkages of the sense strand are chemically modified.
  • At least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are modified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are modified.
  • At least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the antisense strand are modified. In some embodiments, substantially all of the nucleotides of the antisense strand are modified. In some embodiments, all of the nucleotides of the antisense strand are modified.
  • At least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense and/or antisense strand are modified.
  • at least 50% of the nucleotides of the antisense strand may be modified.
  • at least 55% of the nucleotides of the antisense strand may be modified.
  • at least 60% of the nucleotides of the antisense strand may be modified.
  • at least 65% of the nucleotides of the antisense strand may be modified.
  • At least 70% of the nucleotides of the antisense strand may be modified.
  • at least 75% of the nucleotides of the antisense strand may be modified.
  • at least 80% of the nucleotides of the antisense strand may be modified.
  • at least 85% of the nucleotides of the antisense strand may be modified.
  • at least 90% of the nucleotides of the antisense strand may be modified.
  • at least 90% of the nucleotides of the antisense strand may be modified.
  • at least 95% of the nucleotides of the antisense strand may be modified.
  • substantially all (or all) of the nucleotides in the antisense strand are modified.
  • at least one of the modified nucleotides comprises a modified sugar’ (e.g., ribose moiety).
  • at least one of the modified nucleotides comprises a modified nucleobase.
  • the sense strand comprises at least one modified intemucleoside linkage and/or the antisense strand comprises at least one modified internucleoside linkage.
  • nucleotides of the of the antisense strand are unmodified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the antisense strand are unmodified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are unmodified.
  • At least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are unmodified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are unmodified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
  • nucleotides of the antisense strand are unmodified. In some embodiments, substantially all of the nucleotides of the antisense strand are unmodified. In some embodiments, all of the nucleotides of the antisense strand are unmodified. [00418] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the antisense strand are modified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the antisense strand are modified.
  • not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are modified.
  • antisense strand comprises one or more non-naturally intemucleoside linkage.
  • at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the internucleoside linkages of the antisense strand arc non-naturally occurring.
  • at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the or internucleoside linkages of the antisense strand are chemically modified.
  • At least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are modified.
  • At least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, substantially all of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, all of the nucleotides of the sense strand and antisense strand are modified.
  • At least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense and/or sense strand and antisense strand are modified.
  • at least 50% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 55% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 60% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 65% of the nucleotides of the sense strand and antisense strand may be modified.
  • At least 70% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 75% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 80% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 85% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 90% of the nucleotides of the sense strand and antisense strand may be modified.
  • at least 90% of the nucleotides of the sense strand and antisense strand may be modified.
  • At least 95% of the nucleotides of the sense strand and antisense strand may be modified. In some embodiments, substantially all (or all) of the nucleotides in the sense strand and antisense strand are modified. In some embodiments, at least one of the modified nucleotides comprises a modified sugar (e.g., ribose moiety). In some embodiments, at least one of the modified nucleotides comprises a modified nucleobase. In some embodiments, the sense strand comprises at least one modified intemucleoside linkage and/or the sense strand and antisense strand comprises at least one modified intemucleoside linkage.
  • not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the sense strand and antisense strand are unmodified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are unmodified.
  • At least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are unmodified.
  • At least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, substantially all of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, all of the nucleotides of the sense strand and antisense strand are unmodified.
  • not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the sense strand and antisense strand are modified.
  • not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand and antisense strand are modified.
  • not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are modified.
  • sense strand and antisense strand comprises one or more non- naturally intemucleoside linkage.
  • at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the internucleoside linkages of the sense strand and antisense strand are non-naturally occurring.
  • at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the or intemucleoside linkages of the sense strand and antisense strand are chemically modified.
  • the RNA agent ⁇ e.g., described herein) ⁇ e.g., RNAi agent, dsRNA agent, ssRNA agent, antisense strand, sense strand, antisense oligonucleotide
  • RNA agent comprises a heterologous moiety ⁇ e.g., operably connected to the RNA agent.
  • conjugates comprising an RNA agent ⁇ e.g., described herein) ⁇ e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) and a heterologous moiety ⁇ e.g., operably connected to the RNA agent).
  • the conjugate can comprise a modified agent ⁇ e.g., described herein, see, e.g., ⁇ 4.3).
  • the heterologous moiety modifies one or more properly of the RNA agent ⁇ e.g., described herein) ⁇ e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) to which it is conjugated.
  • RNA agent e.g., described herein
  • exemplary properties include, but are not limited to, pharmacodynamics, pharmacokinetics, stability, absorption, activity, tissue distribution, cellular distribution, cellular uptake, charge, half-life, clearance, and binding affinity to a target nucleic acid molecule ⁇ e.g., a target mRNA).
  • the heterologous moiety enhances the distribution and/or uptake ⁇ e.g., into a cell, e.g., into a cell in a subject) of the RNA agent ⁇ e.g., described herein) ⁇ e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) ⁇ e.g., as compared to an RNA agent that lacks the heterologous moiety).
  • the heterologous moiety alters ⁇ e.g., extends) the lifetime ⁇ e.g., in vivo) of the RNA agent ⁇ e.g., described herein) ⁇ e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) ⁇ e.g., as compared to an RNA agent that lacks the heterologous moiety).
  • the heterologous moiety provides an enhanced affinity for a selected target, e.g., a selected molecule, cell type, compartment ⁇ e.g., cell type, tissue, organ or region of the body) ⁇ e.g., as compared to an RNA agent that lacks the heterologous moiety).
  • the heterologous moiety enhances the activity ⁇ e.g., in a cell, e.g., in a cell in a subject) of the RNA agent ⁇ e.g., described herein) ⁇ e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) ⁇ e.g., as compared to an RNA agent that lacks the heterologous moiety).
  • Activity can include, e.g., degradation of a target mRNA ⁇ e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), inhibition of expression of a target gene ⁇ e.g., a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 gene), and/or reduction in the expression of a target gene ⁇ e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 gene).
  • the heterologous moiety imparts a new property on the RNA agent ⁇ e.g., described herein) ⁇ e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) to which it is conjugated.
  • RNA agent e.g., described herein
  • fluorophores or reporter groups that enable detection of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
  • heterologous moieties can impart multiple (e.g., any combination of the foregoing) properties of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
  • RNA agent e.g., described herein
  • dsRNA agent e.g., RNAi agent, dsRNA agent, antisense strand, sense strand
  • the heterologous moiety does not take part in, does not alter, and/or does not interfere with, the creation of a double strand region.
  • Heterologous moieties include for example, but are not limited to, carbohydrates, peptides, proteins (e.g., antibodies or functional fragments or variants thereof; ligands (e.g., of a target receptor)), lipids, polymers, small molecules, intcrcalators, reporter molecules, polyamincs, polyamides, vitamin moieties, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipophilic groups, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores, and dyes.
  • the heterologous moiety is a carbohydrate, peptide, protein (e.g., antibody or functional fragment or variant thereof, e.g., ligand (e.g., of a target receptor)), lipid, polymer, small molecule, or any combination thereof.
  • the heterologous moiety comprises an active drug substance. In some embodiments, the heterologous moiety does not contain an active drug substance.
  • the heterologous moiety is a targeting moiety.
  • the targeting moiety enhances distribution of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) to a target cell (or population of cells), tissue, and/or organ (e.g., as compared to an RNA agent that lacks the targeting moiety).
  • the targeting moiety enhances the uptake of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) into a target cell (or population of cells) (e.g., as compared to an RNA agent that lacks the targeting moiety).
  • the targeting moiety provides enhanced affinity for a selected target, e.g., molecule, cell, cell type, compartment, e.g., a cellular or organ compartment, tissue, organ or region of the body, (e.g., as compared to an RNA agent that lacks the targeting moiety).
  • the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed on the surface of a target cell, tissue, and/or organ.
  • a target molecule e.g., protein, carbohydrate, lipid, etc.
  • the target molecule is a protein, carbohydrate, or lipid.
  • the target molecule is a receptor.
  • the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by one or more population of immune cells (e.g., on the surface of the surface of one or more population of immune cells).
  • a target molecule protein e.g., receptor
  • the immune cells are within the microenvironment of a tumor. In some embodiments, the immune cells are characterized as tumor infiltrating lymphocytes.
  • the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by T-cells (e.g., on the surface of the surface of T- cells).
  • a target molecule protein e.g., receptor
  • the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by CD4+ T-cells (e.g., on the surface of the surface of CD4+ T-cells).
  • the targeting moiety specifically binds to a target molecule protein (e.g., receptor) expressed on the surface of CD4+ T-cells. In some embodiments, the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by CD8+ T-cells (e.g., on the surface of the surface of CD8+ T-cells). In some embodiments, the targeting moiety specifically binds to a target molecule protein (e.g., receptor) expressed on the surface of CD8+ T-cells.
  • a target molecule protein e.g., receptor
  • the T-cells are within the microenvironment of a tumor. In some embodiments, the T-cells are characterized as tumor infiltrating T-cells. In some embodiments, the CD4+ T-cells are within the microenvironment of a tumor. In some embodiments, the CD4+ T- cells are characterized as tumor infiltrating CD4+ T-cells. In some embodiments, the CD8+ T- cells are within the microenvironment of a tumor. In some embodiments, the CD8+ T-cells are characterized as tumor infiltrating CD8+ T-cells.
  • the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by B-cells (e.g., on the surface of the surface of B- cells).
  • a target molecule protein e.g., receptor
  • the B-cells are within the microenvironment of a tumor.
  • the B-cells are characterized as tumor infiltrating B-cells.
  • the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) is directly attached to the heterologous moiety (e.g., targeting moiety) (e.g., directly attached through a single chemical bond).
  • the RNA agent e.g., described herein
  • the RNA agent e.g., RNAi agent, dsRNA agent, antisense strand, sense strand
  • the heterologous moiety e.g., targeting moiety
  • the RNA agent e.g., described herein
  • the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) is indirectly attached to the heterologous moiety via a linker.
  • Suitable linkers for use in the conjugates described herein are known in the art and can be evaluated by a person of ordinary skill in the ait using standard methods. Exemplary linkers and components thereof for use in the conjugates described herein are also described below.
  • Linkers typically comprise a direct bond or an atom such as oxygen or sulfur, a unit such as NR8, C(O), C(O)NH, SO, SO2, SO2NH or a chain of atoms, such as, but not limited to, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl, alkeny
  • alkynylheteroarylalkenyl alkynylheteroarylalkynyl, alkylheterocyclylalkyl alkylheterocyclylalkenyl, alkylhererocyclylalkynyl, alkenylheterocyclylalkyl. alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl.
  • alkynylheterocyclyl alkenyl, alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl, alkylhctcroaryl, alkcnylhctcroaryl, alkynylhcrcroaryl, which one or more methylenes can be inteiTupted or terminated by O, S, S(O), SO2, N(R8), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocyclic; where R8 is hydrogen, acyl, aliphatic, or substituted aliphatic.
  • the linker is about 1-24 atoms, 2-24, 3-24, 4-24, 5-24, 6-24, 6-18, 7-18, 8-18, 7-17, 8-17, 6-16, 7-17, or 8-16 atoms.
  • the linker comprises ethylene glycol, nucleosides, or amino acid units. In some embodiments, the linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In some embodiments, the linker comprises groups selected from alkyl, amino, oxo, amide and ether groups. In some embodiments, the linker comprises groups selected from alkyl and amide groups. In some embodiments, the linker comprises groups selected from alkyl and ether groups. In some embodiments, the linker comprises at least one phosphorus moiety. In some embodiments, the linker comprises at least one phosphate group.
  • the linker comprises at least one neutral linking group.
  • exemplary linkers include but are not limited to pyrrolidine, 8-amino- 3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC), 6-aminohexanoic acid (AHEX or AHA).
  • Additional exemplary linkers include but are not limited to substituted or unsubstituted Ci-Cw alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted C2-C10 alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.
  • the linker is bifunctional.
  • a bifunctional linker comprises at least two functional groups. One of the functional groups is selected to react with a particular' site on an RNA agent (e.g., described herein) and the other is selected to react with a heterologous moiety (e.g., described herein).
  • Examples of functional groups used in a bifunctional linkers include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups.
  • bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.
  • the linker is a monovalent linker, a bivalent linker, a trivalent linker, or a tetravalent linker.
  • the linker is non-cleavable. In some embodiments, the linker is cleavable. Cleavable linkers contain at least one (or a plurality of) cleavable bonds that are susceptible to one or more cleavage agent. Exemplary classes of cleavable linkers include, but are not limited to, redox clcavablc linkers, phosphate based clcavablc linkers, acid clcavablc linkers, ester-based cleavable linkers, and peptide-based cleavable linkers.
  • a cleavable bond is selected from among: an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, or a disulfide.
  • Cleavable linkers may be advantageous when a stable conjugate is desired under a first set of conditions but under a second set of conditions it is advantageous to release the RNA agent (e.g., described herein) from the heterologous moiety (e.g., described herein).
  • RNA agent e.g., described herein
  • the linker is not cleaved (or is cleaved at a lower rate) under a first condition relative to under a second condition.
  • the first condition is within the blood (e.g., of a subject) (or in an in vitro system sufficient to mimic the conditions of the blood within a subject) and the second condition is with a cell (e.g., a cell within a subject) (or in an in vitro system sufficient to mimic the conditions of a cell within a subject).
  • the suitability of a cleavable linker can be assessed by standard methods known in the art. In general, the suitability of a cleavable linker can be evaluated by testing the ability of a cleavage agent (or condition) to cleave the candidate linker (e.g., the cleavage bond(s)). In some embodiments, it may be desirable to further test the ability of the linker to resist cleavage under a certain condition (e.g., within the blood or serum of subject, when in contact with a non-target cell, tissue, organ).
  • a cleavage agent or condition
  • it may be desirable to further test the ability of the linker to resist cleavage under a certain condition e.g., within the blood or serum of subject, when in contact with a non-target cell, tissue, organ.
  • the linker is a redox cleavable linker that is cleaved upon reduction or oxidation.
  • a reductively cleavable linker is a disulphide (-S-S-) containing linker.
  • Redox cleavable linkers can be evaluated using methods analogous to those described above.
  • the linker is a phosphate-based cleavable linker.
  • a phosphate- based cleavable linker is cleaved by agents that degrade or hydrolyze the phosphate group.
  • agents that degrade or hydrolyze the phosphate group For example, in cells, enzymes such as phosphatases arc capable of cleaving phosphate groups.
  • phosphate -based linkers include those comprising any of the following -O- P(O)(ORk)-O-, -OP(S)(ORk)-O-, -O-P(S)(SRk)-O-, -S-P(O)(ORk)-O-, -O-P(O)(ORk)-S-, -S- P(O)(ORk)-S-, -OP(S)(ORk)-S-, -S-P(S)(ORk)-O-, -O-P(O)(Rk)-O-, -O-P(S)(Rk)-O-, -S- P(O)(Rk)-O-, -S-P(S)(Rk)-O-, -S-P(S)(Rk)-O-, -S-P(O)(Rk)-S-, -O-P(S)( Rk)-S-, wherein
  • Exemplary embodiments include are -OP(O)(OH)-O-, -O-P(S)(OH)-O-, -O-P(S)(SH)-O-, -S- P(O)(OH)-O-, -O-P(O)(OH)-S-, -S-P(O)(OH)-S-, -O-P(S)(OH)-S-, -S-P(S)(OH)-O-, -O-P(O)(H)- O-, -O-P(S)(H)-O-, -S-P(O)(H)-O, -S-P(O)(H)-O-, -S-P(O)(H)-S-, or -O-P(S)(H)-S-.
  • Phosphate based cleavable linker can be evaluated using methods analogous to those described above.
  • the linker is an acid cleavable linker.
  • An acid cleavable linker is cleaved under acidic conditions.
  • the acid cleavable linker can be cleaved in an acidic environment with a pH of about 6.5 or less (e.g., about 6.0, 5.5, 5.0, or less).
  • the acid cleavable linker can be cleaved by enzymes that can act as a general acid.
  • specific low pH organelles such as endosomes and lysosomes can provide a cleaving environment for acid cleavable linkers.
  • acid cleavable linkers include but are not limited to hydrazones, esters, and esters of amino acids.
  • Acid cleavable linkers can be evaluated using methods analogous to those described above.
  • the linker is an ester-based cleavable linker.
  • An ester-based cleavable linker is cleaved by enzymes such as esterases and amidases in cells.
  • Examples of ester- based cleavable include, but are not limited to, esters of alkylene, alkenylene and alkynylene groups.
  • the cleavable bonds of ester cleavable linkers have the general formula -C(O)O- or - OC(O)-. Ester-based cleavable linkers can be evaluated using methods analogous to those described above.
  • the linker is a peptide-based cleavable linker.
  • a peptide-based cleavable linker is cleaved by enzymes such as peptidases and proteases (e.g., present in cells (e.g., cells within a subject)).
  • Peptide-based cleavable linkers comprise peptide bonds formed between amino acids to yield polypeptides (e.g., dipeptides, tripeptides, etc.). As known in the art, peptide bonds.
  • the peptide bonds (i.e., the amide bond) of the peptide linker is generally the site of cleavage.
  • Peptide-based cleavable linkers can be evaluated using methods analogous to those described above.
  • the heterologous moiety may be attached at any suitable position to the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
  • RNA agent e.g., described herein
  • RNAi agent e.g., RNAi agent, dsRNA agent, antisense strand, sense strand.
  • the heterologous moiety is conjugated to the 5' end of the RNA agent (e.g., described herein) e.g., RNAi agent, dsRNA agent, antisense strand, sense strand). In some embodiments, the heterologous moiety is conjugated to the 3' end of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
  • a first heterologous moiety is conjugated to the 5' end of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) and a second heterologous moiety is conjugated to the 3' end of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
  • the first and second heterologous moieties can be the same or different.
  • the heterologous moiety is conjugated to an internal site of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
  • the RNA agent (e.g., RNAi agent, dsRNA agent) comprises an antisense strand.
  • the heterologous moiety is conjugated to the 5' end of the antisense strand.
  • the heterologous moiety is conjugated to the 3' end of the antisense strand.
  • a first heterologous moiety is conjugated to the 5' end of the antisense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand.
  • the first and second heterologous moieties can be the same or different.
  • the heterologous moiety is conjugated to an internal site of the antisense strand.
  • the heterologous moiety may be attached to the 3' end of the sense and/or antisense strand.
  • the heterologous moiety may be attached to the 5' end of the sense and/or antisense strand.
  • the heterologous moiety may be attached to at an internal site of the sense and/or antisense strand.
  • the heterologous moiety may be attached to the 3' end of the sense and antisense strand.
  • the heterologous moiety may be attached to the 5' end of the sense and antisense strand.
  • the heterologous moiety may be attached to at an internal site of the sense and antisense strand.
  • the RNA agent (e.g., RNAi agent, dsRNA agent) comprises a sense strand.
  • the heterologous moiety is conjugated to the 5' end of the sense strand.
  • the heterologous moiety is conjugated to the 3' end of the sense strand.
  • a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the sense strand.
  • the first and second heterologous moieties can be the same or different.
  • the heterologous moiety is conjugated to an internal site of the sense strand.
  • the RNA agent (e.g., RNAi agent) comprises a dsRNA agent comprising a sense strand and an antisense strand
  • the heterologous moiety is conjugated to the 5' end of the sense strand.
  • the heterologous moiety is conjugated to the 3' end of the sense strand.
  • a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the sense strand.
  • the first and second heterologous moieties can be the same or different.
  • the heterologous moiety is conjugated to an internal site of the sense strand.
  • the heterologous moiety is conjugated to the 5' end of the sense strand. In some embodiments, the heterologous moiety is conjugated to the 3' end of the sense strand. In some embodiments, a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the sense strand. The first and second heterologous moieties can be the same or different. In some embodiments, the heterologous moiety is conjugated to an internal site of the sense strand.
  • a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand.
  • a first heterologous moiety is conjugated to the 3' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand.
  • a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand.
  • a first heterologous moiety is conjugated to the 3' end of the sense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand.
  • the first and second heterologous moieties can be the same or different.
  • a first heterologous moiety is conjugated to an internal site of the sense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to an internal site of the sense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to an internal site of the antisense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand.
  • a first heterologous moiety is conjugated to an internal site of the antisense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand.
  • the first and second heterologous moieties can be the same or different.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • inhibits expression of a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 in a cell.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent e.g., RNAi agent, e.g., dsRNA agent
  • RNAi agent e.g., dsRNA agent
  • a target gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6
  • a subject e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.
  • the RNA agent mediates degradation of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 (e.g., hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6) mRNA).
  • a target mRNA e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 (e.g., hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6) mRNA.
  • KLHL6 e.g., hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6
  • PCR e.g., qPCR
  • branched DNA assays or by a protein-based methods (such as immunofluorescence analysis (using, e.g., western blotting or flow cytometric techniques).
  • a protein-based methods such as immunofluorescence analysis (using, e.g., western blotting or flow cytometric techniques).
  • inhibition of gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 (e.g., hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6)
  • KLHL6 e.g., hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6
  • the cell is an immune cell.
  • immune cells include, T cells (e.g., CD4+ T cells, CD8+ T cells), B-cells, tumor infiltrating lymphocytes (e.g., tumor infiltrating T cells (e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells)).
  • T cells e.g., CD4+ T cells, CD8+ T cells
  • B-cells tumor infiltrating lymphocytes
  • tumor infiltrating T cells e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells
  • the cell is a T-cell.
  • the cell is a tumor infiltrating lymphocyte.
  • the cell is a tumor infiltrating T cell.
  • the cell is a tumor infiltrating CD4+ T cell.
  • the cell is a CD8+ tumor infiltrating T cell.
  • the cell is B-cell.
  • RNA agent e.g., described herein
  • RNAi agent e.g., RNAi agent, dsRNA agent, antisense strand, sense strand
  • chemical synthesis e.g., solid phase synthesis
  • RNA agent described herein
  • RNAi agent e.g., RNAi agent, dsRNA agent, antisense strand, sense strand
  • single stranded nucleic acid molecules e.g., described herein
  • dsRNA agents e.g., described herein
  • dsRNA agents can be prepared using a two-step procedure, wherein the individual strands of the dsRNA agent are prepared separately and subsequently annealed.
  • the individual strands of the dsRNA agent can be prepared using solutionphase or solid-phase organic synthesis or both.
  • the RNA agents e.g., described herein
  • a solution e.g., an aqueous or organic solution
  • the dsRNA agent can be precipitated and redissolved in pure double-distilled water, and lyophilized.
  • the lyophilized dsRNA agent can be resuspended in a solution appropriate for the intended formulation process.
  • conjugates can be synthesized utilizing standard methods known in the ait. See, e.g., Dong Y, Siegwart DJ, Anderson DG. Strategies, design, and chemistry in siRNA delivery systems. Adv Drug Deliv Rev. 2019 Apr;144: 133-147. doi: 10.1016/j.addr.2019.05.004. Epub 2019 May 15. PMID: 31102606; PMCID: PMC6745264, the entire contents of which is incorporated herein by reference for all purposes.
  • standard conjugation methods include, e.g., parallel synthesis methods and linear synthesis methods.
  • RNA agents described herein e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands
  • a vector e.g., a non-viral vector (e.g., a plasmid), a viral vector).
  • vectors e.g., non-viral vectors (e.g., plasmids) viral vectors) comprising one or more agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand) (see, e.g., ⁇ 4.2, 4.3).
  • agent described herein e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand
  • agent described herein e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand
  • the vector used can be any vector that is suitable for cloning nucleic acid molecules that can be used for transcription of the nucleic acid molecule of interest (e.g., an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand) (see, e.g., ⁇ 4.2, 4.3).
  • RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand) (see, e.g., ⁇ 4.2, 4.3).
  • the vector is a viral vector.
  • Viral vectors include both RNA and DNA based vectors.
  • the vectors can be designed to meet a variety of specifications.
  • viral vectors can be engineered to be capable or incapable of replication in prokaryotic and/or eukaryotic cells.
  • the vector is replication deficient.
  • the vector is replication competent. Vectors can be engineered or selected that either will (or will not) integrate in whole or in part into the genome of host cells, resulting (or not (e.g., episomal expression)) in stable host cells comprising the desired nucleic acid in their genome.
  • Exemplary viral vectors include, but are not limited to, adenovirus vectors, adeno- associated virus vectors, lentivirus vectors, retrovirus vectors, poxvirus vectors, parapoxivirus vectors, vaccinia virus vectors, fowlpox virus vectors, herpes virus vectors, adeno-associated virus vectors, alphavirus vectors, lentivirus vectors, rhabdovirus vectors, measles virus, Newcastle disease virus vectors, picornaviruses vectors, anellovectors, or lymphocytic choriomeningitis virus vectors.
  • the viral vector is an adenovirus vector, adeno-associated virus vector, lentivirus vector, anellovector (as described, for example, in U.S. Pat. No. 11,446,344, the entire contents of which is incorporated by reference herein for all purposes).
  • the vector is an adenoviral vector (e.g., human adenoviral vector, e.g., HAdV or AdHu).
  • the adenovirus vector has the El region deleted, rendering it replication-deficient in human cells. Other regions of the adenovirus such as E3 and E4 may also be deleted.
  • Exemplary adenovirus vectors include, but are not limited to, those described in e.g., W02005071093 or WQ2006048215, the entire contents of each of which is incorporated by reference herein for all purposes.
  • the adenovirus-based vector used is a simian adenovirus, thereby avoiding dampening of the immune response after vaccination by pre-existing antibodies to common human entities such as AdHu5.
  • simian adenovirus vectors include AdCh63 (see, e.g., W02005071093, the entire contents of which is incorporated by reference herein for all purposes) or AdCh68.
  • Viral vectors can be generated through the use of a packaging/producer cell line (e.g., a mammalian cell line) using standard methods known to the person of ordinary skill in the art.
  • a nucleic acid construct e.g., a plasmid
  • the transgene e.g., an RNA agent described herein
  • additional elements e.g., a promoter, inverted terminal repeats (ITRs) flanking the transgene
  • a plasmid encoding e.g., viral replication and structural proteins along with one or more helper plasmids
  • a host cell e.g., a host cell line
  • a host cell line i.e., the packing/producer cell line
  • helper plasmid may also be needed that include helper genes from another virus (e.g., in the instance of adeno-associated viral vectors).
  • Eukaryotic expression plasmids are commercially available from a variety of suppliers, for example the plasmid series: pcDNATM, pCR3.1 TM, pCMVTM, pFRTTM, pVAXl TM, pCITM, NanoplasmidTM, and Pcaggs.
  • the person of ordinary skill in the art is aware of numerous transfection methods and any suitable method of transfection may be employed (e.g., using a biochemical substance as carrier (e.g., lipofectamine), by mechanical means, or by electroporation,).
  • the cells are cultured under conditions suitable and for a sufficient time for plasmid expression.
  • the viral particles may be purified from the cell culture medium using standard methods known to the person of ordinary skill in the art. For example, by centrifugation followed by e.g., chromatography or ultrafiltration.
  • the vector is a plasmid.
  • a person of ordinary skill in the art is aware of suitable plasmids for expression of the DNA of interest.
  • Suitable plasmid DNA may be generated to allow efficient production of the encoded RNA in cell lines, e.g., in insect cell lines, for example using vectors as described in W02009150222A2 and as defined in PCT claims 1 to 33, the disclosure relating to claim 1 to 33 of W02009150222A2 the entire contents of which is incorporated by reference herein for all purposes.
  • RNA agent described herein e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand
  • a vector described herein see, e.g., ⁇ 4.7
  • a conjugate described herein see, e.g., ⁇ 4.4
  • a carrier described herein see, e.g., ⁇ 4.9
  • a pharmaceutical composition described herein see, e.g., ⁇ 4.10), and/or a cell described herein (see, e.g., ⁇ 4.8); or any combination thereof.
  • the cell or population of cells is in vitro. In some embodiments, the cell or population of cells is in vivo. In some embodiments, the cell or population of cells is ex vivo. In some embodiments, the cell or population of cells are derived from a subject (e.g., a human subject. In some embodiments, the subject is a human, mouse, rat, primate, or non-human primate.
  • a subject e.g., a human subject. In some embodiments, the subject is a human, mouse, rat, primate, or non-human primate.
  • the cell (or population of cells) is an immune cell. In some embodiments, the cell (or population of cells) is a subset of immune cells.
  • the immune cells are within the microenvironment of a tumor. In some embodiments, the immune cells are characterized as tumor infiltrating lymphocytes.
  • the cell (or population of cells) is a CD4+ T cell. In some embodiments, the cell (or population of cells) is a CD8+ T cell. In some embodiments, the population of cells comprises is CD8+ T cells and CD4+ T cells.
  • the T-cells are within the microenvironment of a tumor. In some embodiments, the T-cells are characterized as tumor infiltrating T-cells. In some embodiments, the CD4+ T-cells are within the microenvironment of a tumor. In some embodiments, the CD4+ T- cells are characterized as tumor infiltrating CD4+ T-cells. In some embodiments, the CD8+ T- cells are within the microenvironment of a tumor. In some embodiments, the CD8+ T-cells are characterized as tumor infiltrating CD8+ T-cells.
  • the cell (or population of cells) is a B cell. In some embodiments, the cell (or population of cells) is a B cell. In some embodiments, the population of cells comprises is B cells.
  • the B-cells are within the microenvironment of a tumor. In some embodiments, the B-cells are characterized as tumor infiltrating B-cells.
  • RNA agents described herein e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands (or a conjugate comprising the same)
  • a vector comprising any of the foregoing is formulated within one or more carrier.
  • carriers comprising any one or more of the RNA agents described herein (e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands (or a conjugate comprising the same)) or a vector comprising any of the foregoing.
  • RNA agents described herein e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands (or a conjugate comprising the same)
  • a vector comprising any of the foregoing can be encapsulated within a carrier, chemically conjugated to a carrier, associated with the carrier.
  • RNA agent described herein (or a conjugate comprising the same) (or a vector comprising the same) with one or more molecules of a carrier (e.g., one or more lipids of a lipid-based carrier, e.g., an LNP, liposome, lipoplex, and/or nanoliposome) into larger complexes or assemblies without covalent binding.
  • a carrier e.g., one or more lipids of a lipid-based carrier, e.g., an LNP, liposome, lipoplex, and/or nanoliposome
  • the term “encapsulation” refers to the incorporation of an RNA agent described herein (or a conjugate comprising the same) (or a vector comprising the same) into a carrier (e.g., a lipid-based carrier, e.g., an LNP, liposome, lipoplex, and/or nanoliposome) wherein the RNA agent described herein (or the conjugate comprising the same) (or the vector comprising the same) is entirely contained within the interior space of the carrier (e.g., the lipid-based carrier, e.g., the LNP, liposome, lipoplex, and/or nanoliposome).
  • a carrier e.g., a lipid-based carrier, e.g., an LNP, liposome, lipoplex, and/or nanoliposome
  • Exemplary carriers includes, but are not limited to, lipid-based carriers (e.g., lipid nanoparticles (LNPs), liposomes, lipoplexes, and nanoliposomes).
  • the carrier is a lipid-based carrier.
  • the carrier is an LNP.
  • the LNP comprises a cationic lipid, a neutral lipid, a cholesterol, and/or a PEG lipid. Lipid based carriers are further described below in ⁇ 4.9.1.
  • an RNA agent described herein e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)
  • a vector comprising any of the foregoing is encapsulated or associated with one or more lipids (e.g., cationic lipids and/or neutral lipids), thereby forming lipid-based carriers such as lipid nanoparticles (LNPs), liposomes, lipoplexes, or nanoliposomes.
  • lipids e.g., cationic lipids and/or neutral lipids
  • an RNA agent described herein e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)
  • a vector comprising any of the foregoing is encapsulated in one or more lipids (e.g., cationic lipids and/or neutral lipids), thereby forming lipid-based earners such as lipid nanoparticles (LNPs), liposomes, lipoplexes, or nanoliposomes.
  • lipids e.g., cationic lipids and/or neutral lipids
  • an RNA agent described herein e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)
  • a vector comprising any of the foregoing is associated with one or more lipids (e.g., cationic lipids and/or neutral lipids), thereby forming lipid-based carriers such as lipid nanoparticles (LNPs), liposomes, lipoplexes, or nanoliposomes.
  • lipids e.g., cationic lipids and/or neutral lipids

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Abstract

Provided herein are, inter alia, agents (e.g., RNAi agents, dsRNA agents) comprising a sense strand and an antisense strand targeting one or more genes expressed by one or more subsets of immune cells, e.g., T cells (e.g., tumor infiltrating T-cells), B-cells; and methods of manufacturing and pharmaceutical compositions comprising the same. Further provided herein are methods of utilizing the RNA agents (e.g., RNAi agents, dsRNA agents) including, e.g., methods of inhibiting or decreasing expression of one or more genes expressed by one or more subsets of immune cells, e.g., T cells (e.g., tumor infiltrating T-cells), B-cells expression (e.g., mRNA expression), methods of treating proinflammatory (e.g., autoimmune) diseases.

Description

IMMUNE CELL TARGETED COMPOSITIONS AND RELATED METHODS
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Serial No.: 63/632,335, filed April 10, 2024; U.S. Serial No.: 63/634,206, filed April 15, 2024; U.S. Serial No.: 63/636,512, filed April 19, 2024; U.S. Serial No.: 63/637,446, filed April 23, 2024; U.S. Serial No.: 63/638,958, filed April 26, 2024; the entire contents of each of which is incorporated herein by reference.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on April 4, 2025, is named 62801_60W001_SL.xml and is 52,915 bytes in size.
1. FIELD
[0003] This disclosure relates to agents (e.g., RNA agents (e.g., double stranded RNAi agents comprising a sense strand and an antisense strand)) targeting one or more genes expressed by one or more subsets of immune cells, e.g., T cells, B-cells. The disclosure further relates to pharmaceutical compositions comprising the same; and methods of utilizing the same, including, e.g., methods of inhibiting expression of one or more target gene expressed by one or more subsets of immune cells e.g., T cells, B-cells, etc.), and methods of treating diseases associated with expression of one or more of said genes (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer) comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.).
2. BACKGROUND
[0004] Somatic mutations are nucleotide alterations in a genomic DNA sequence (e.g., of a gene) that are acquired during the lifetime of a subject. Unlike germline mutations, somatic mutations are not present in the germline DNA of an individual and are therefore not inherited from a parent like germline polymorphisms. Somatic mutations may occur spontaneously due to e.g., the infidelity of DNA replication occurring at each cell division creating substitutions, deletions, or additions of nucleotides into the DNA of a cell. A somatic mutation may also be caused by environmental factors such as e.g., ultraviolet radiation, chemical exposure, or virial infections.
3. SUMMARY
[0005] Provided herein are, inter alia, agents (e.g., RNA agents (e.g., RNAi agents, dsRNA agents, ssRNA agents, siRNA agents, antisense oligonucleotides, etc.)) targeting one or more genes expressed by one or more subsets of immune cells, e.g., T cells, B-cells, tumor infiltrating lymphocytes (TILs) (e.g., tumor infiltrating T-cells); and methods of manufacturing and pharmaceutical compositions comprising the same. Further provided herein are methods of utilizing the agents (e.g., RNA agents (e.g., RNAi agents, dsRNA agents, etc.)) including, e.g., methods of inhibiting or decreasing gene expression (e.g., mRNA expression), methods of treating diseases associated with the one or more genes, and methods of treating diseases, including, e.g., proinflammatory diseases (e.g., autoimmune diseases); and cancer; and methods of treating diseases associated with expression of one or more of said genes (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer) comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.).
[0006] Accordingly, in one aspect, provided herein are double stranded ribonucleic acid (dsRNA) agents for inhibiting expression of BTG1 (e.g., human BTG1 (hBTGl)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
[0007] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
[0008] In one aspect, provided herein are dsRNA agents for inhibiting expression of BTG1 (e.g., (hBTGl) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding BTG1 (e.g., (hBTGl).
[0009] In some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding BTG1 (e.g., (hBTG1 ). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
[0010] In one aspect, provided herein are dsRNA agents for inhibiting expression of BTG1 (e.g., (hBTGl) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding BTG1 (e.g., (hBTGl), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
[0011] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
[0012] In one aspect, provided herein are double stranded ribonucleic (dsRNA) agents for inhibiting expression of CDKN1B (e.g., human CDKN1B (hCDKNIB)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
[0013] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
[0014] In one aspect, provided herein arc dsRNA agents for inhibiting expression of CDKN1B (e.g., (hCDKNIB) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding CDKN1B (e.g., (hCDKNIB).
[0015] In some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding CDKN1B (e.g., (hCDKNIB). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
[0016] In one aspect, provided herein are dsRNA agents for inhibiting expression of CDKN1B (e.g., (hCDKNIB) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding CDKN1B (e.g., (hCDKNIB), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
[0017] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
[0018] In one aspect, provided herein are double stranded ribonucleic (dsRNA) agents for inhibiting expression of DUSP2 (e.g., human DUSP2 (hDUSP2)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
[0019] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e. ., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
[0020] In one aspect, provided herein are dsRNA agents for inhibiting expression of DUSP2 (e.g., (hDUSP2) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding DUSP2 (e.g., (hDUSP2), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
[0021] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
[0022] In one aspect, provided herein are dsRNA agents for inhibiting expression of DUSP2 (e.g., (hDUSP2) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding DUSP2 (e.g., (hDUSP2).
[0023] In some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding DUSP2 (e.g., (hDUSP2)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
[0024] In one aspect, provided herein are dsRNA agents for inhibiting expression of DUSP2 (e.g., (hDUSP2) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding DUSP2 (e.g., (hDUSP2), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
[0025] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
[0026] In one aspect, provided herein are double stranded ribonucleic (dsRNA) agents for inhibiting expression of IGLL5 (e.g., human IGLL5 (hIGLL5)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
[0027] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
[0028] In one aspect, provided herein are dsRNA agents for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding IGLL5 (e.g., (hIGLL5).
[0029] In some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 (e.g., (hIGLL5)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
[0030] In one aspect, provided herein are dsRNA agents for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
[0031] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
[0032] In one aspect, provided herein are double stranded ribonucleic (dsRNA) agents for inhibiting expression of IGLL5 (e.g., human IGLL5 (hIGLL5)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
[0033] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
[0034] In one aspect, provided herein arc dsRNA agents for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding IGLL5 e.g., (hIGLL5).
[0035] In some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 e.g., (hIGLL5)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
[0036] In one aspect, provided herein are dsRNA agents for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA sequence encoding IGLL5 (e.g., (hIGLL5), wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 14.
[0037] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region sequence of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
[0038] In one aspect, provided herein are double stranded ribonucleic (dsRNA) agents for inhibiting expression of KLHL6 (e.g., human KLHL6 (hKLHL6)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
[0039] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
[0040] In one aspect, provided herein are dsRNA agents for inhibiting expression of KLHL6 (e.g., (hKLHL6) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding KLHL6 (e.g., (hKLHL6).
[0041] In some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding KLHL6 (e.g., (hKLHL6)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
[0042] In one aspect, provided herein are dsRNA agents for inhibiting expression of KLHL6 (e.g., (hKLHL6) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding KLHL6 (e.g., (hKLHL6), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
[0043] In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.1n some embodiments, the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 (e.g., (hIGLL5)). In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides. In some embodiments, the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
[0044] For the sake of clarity, it should be understood that the following exemplary embodiments, can be applied to any of the foregoing aspects.
[0045] In some embodiments, the send strand comprises at least one modified nucleotide and/or the antisense strand comprises at least one modified nucleotide.
[0046] In some embodiments, the sense strand comprises at least one modified nucleotide and/or the antisense strand comprises at least one modified nucleotide. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and/or antisense strand are modified. In some embodiments, substantially all (or all) of the nucleotides in the sense strand and/or antisense strand are modified. In some embodiments, at least one of the modified nucleotides comprises a modified sugar (e.g., ribose moiety). In some embodiments, at least one of the modified nucleotides comprises a modified nucleobase. In some embodiments, the sense strand comprises at least one modified internucleoside linkage and/or the antisense strand comprises at least one modified intemucleoside linkage.
[0047] In some embodiments, each of the antisense strand and the sense strand are not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides in length. In some embodiments, the antisense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-30 20- 30, 21-30, 22-30, 23-30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21 , 19-22, 19-23, 19-24, or 19-25 nucleotides; and/or the sense strand comprises from about 15-30, 16-30, 17-30,
18-30, 19-30 20-30, 21-30, 22-30, 23-30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19- 21, 19-22, 19-23, 19-24, or 19-25 nucleotides. In some embodiments, the antisense strand comprises or consists of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides; and/or the antisense strand comprises or consist of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides. In some embodiments, antisense strand comprises from about 19-23 nucleotides; and/or the sense strand comprises from about 19-23 nucleotides. In some embodiments, antisense strand comprises or consists of about 23 nucleotides; and/or the sense strand comprises or consists of about 21 nucleotides.
[0048] In some embodiments, the sense strand and/or the antisense strand comprises a 3' and/or 5' overhang of 1, 2, or 3 nucleotides. In some embodiments, the antisense strand comprises a 3' overhang of 1, 2, or 3 nucleotides (e.g., 2 nucleotides).
[0049] In some embodiments, the double stranded region is from about 19-30, 19-29, 19-28,
19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-20, 19-21, 23-30, 23-29, 23-28, 23-27, 23-26, 23-25, 23-24, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotide pairs in length. In some embodiments, the double stranded region is from about 19-23 or 19-21 nucleotide pairs in length. In some embodiments, the double stranded region is about 21 nucleotide pairs in length. In some embodiments, the sense strand and the antisense strand are part of a single nucleic acid molecule (e.g., wherein a hairpin loop is between the sense strand and the antisense strand of the single nucleic acid molecule). In some embodiments, the sense strand and the antisense strand are separate nucleic acid molecules (i.e., connected only through the double stranded region).
[0050] In one aspect, provided herein are conjugates comprising a dsRNA agent described herein and a heterologous moiety. In some embodiments, the heterologous moiety is a peptide, protein, carbohydrate, lipid, polymer, or small molecule. In some embodiments, the heterologous moiety is attached to the dsRNA agent via a linker. In some embodiments, the heterologous moiety attached to the 3' end of the sense and/or antisense strand and/or the 5' end of the sense and/or antisense strand, and/or at an internal site of the sense and/or antisense strand.
[0051] In one aspect, provided herein are vectors (e.g., a viral vector, a non-viral vector) encoding an antisense strand, a sense strand, or both an antisense and sense strand of an RNA agent (e.g., dsRNA agent) described herein, or a conjugate described herein. [0052] In one aspect, provided herein are carriers comprising a dsRNA agent described herein, a conjugate described herein, or a vector described herein. In some embodiments, the carrier comprises a nanoparticle, a polymer, a lipid-based delivery system, as dendrimer, a cationic delivery system, or a hydrogel. In some embodiments, the lipid-based delivery system is a lipid nanoparticle (LNP), liposome, lipoplex, nanoliposome, an exosome, or a micelle.
[0053] In one aspect, provided herein are cells comprising a dsRNA agent described herein, a conjugate described herein, a vector described herein, or a carrier described herein.
[0054] In one aspect, provided herein are pharmaceutical compositions comprising a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a cell described herein, and a pharmaceutically acceptable excipient.
[0055] In one aspect, provided herein are kits comprising a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein.
[0056] In one aspect, provided herein are diagnostic kits comprising one or more reagent for use in a method of detecting the presence or absence of one or more somatic BTG1 (e.g., hBTGl), CDKN1B, DUSP2, IGLL5, or KLHL6 mutation in a sample from a subject.
[0057] In one aspect, provided herein are methods of treating, ameliorating, or preventing a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in a subject, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and (c) administering to the subject a therapeutic agent if the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
[0058] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
[0059] In some embodiments, the gene associated disease is a BTG1 associated disease, a CDKN1B associated disease, DUSP2 associated disease, a ILL5 associated disease, or a KLHL6 associated disease.
[0060] In some embodiments, (i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[0061] In some embodiments, the therapeutic agent comprises an anti-inflammatory agent (e.g., an anti-inflammatory agent described herein).
[0062] In some embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
[0063] In one aspect, provided herein are methods of treating, ameliorating, or preventing a proinflammatory disease in a subject, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and (c) administering to the subject a therapeutic agent if the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein. [0064] In some embodiments, the gene is BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
[0065] In some embodiments, the therapeutic agent is an anti-inflammatory agent as described herein. In some embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
[0066] In one aspect, provided herein are methods of diagnosing a gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in a subject, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the presence of the one or more somatic mutation (e.g., a somatic mutation set forth in Table 2) indicates that the subject has a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease.
[0067] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein. In some embodiments, the gene is associated disease is a BTG1 associated disease, a CD KN IB associated disease, DUSP2 associated disease, a IGLL5 associated disease, or a KLHL6 associated disease.
[0068] In some embodiments, (i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[0069] In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory (e.g., autoimmune) disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease.
[0070] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA. [0071] In some embodiments, the method further comprises administering to the subject a therapeutic agent if a somatic mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
[0072] In some embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
[0073] In one aspect, provided herein are methods of selecting a subject for administration of a therapeutic agent, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified, from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the subject is selected for administration of the therapeutic agent if the one or more somatic mutation (e.g., a somatic mutation set forth in Table 2) is present. [0074] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA. In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
[0075] In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
[0076] In some embodiments, the method further comprises administering to the subject the therapeutic agent if the subject is selected. In some embodiments, the therapeutic agent is an antiinflammatory agent as described herein. In some embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the inhibitory nucleic acid molecule comprises a dsRNA agent as described herein, a conjugate as described herein, a vector as described herein, a carrier as described herein, a cell as described herein, a pharmaceutical composition as described herein.
[0077] In one aspect, provided herein are in vitro methods of screening a sample from a subject for one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2), the method comprising (a) isolating and purifying DNA, RNA, or protein from a sample obtained from the subject; and (b) detecting the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[0078] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
[0079] In some embodiments, the sample is a blood, tissue, or cell sample. In some embodiments, the sample is biopsy. In some embodiments, the sample comprises a cell (e.g., a T- ccll) or a population of cells (e.g., a population of T-cclls).
[0080] In some embodiments, at least one of the one or more somatic mutation is a loss of function mutation, a gain of function mutation, a missense mutation, synonymous, and/or non- synonymous. In some embodiments, the one or more somatic mutation is a loss of function mutation. In some embodiments, the one or more somatic mutation truncates the protein encoded by the gene. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth herein. In some embodiments, the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
[0081] In one aspect, provided herein are methods of reducing or inhibiting expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell, the method comprising delivering into the cell the a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell.
[0082] In one aspect, provided herein are methods of reducing or inhibiting expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject, the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject.
[0083] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
[0084] In one aspect, provided herein are methods of treating, ameliorating, or preventing a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in a subject, the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby treat, ameliorate, or prevent the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in the subject.
[0085] In some embodiments, the gene associated disease is a BTG1 associated disease, a CDKN1B associated disease, a DUSP2 associated disease, an IGLL5 associated disease, or a KLHL6 associated disease. [0086] In some embodiments, the gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease associated disease is a proinflammatory disease (e.g., an autoimmune disease). [0087] In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease associated disease is an autoimmune disease.
[0088] In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is treated, ameliorated, or prevented through the reduction or inhibition expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
[0089] In one aspect, provided herein are methods of treating, ameliorate, or preventing a proinflammatory (e.g., autoimmune) disease in a subject, the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby treat, ameliorate, or prevent the proinflammatory (e.g., autoimmune) disease in the subject.
[0090] In some embodiments, the proinflammatory (e.g., autoimmune) disease is treated, ameliorated, or prevented through a reduction or inhibition of expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
[0091] In some embodiments, the subject is a human.
4. DETAILED DESCRIPTION
[0092] The inventors have, inter alia, discovered somatic mutations in select genes expressed by subsets of immune cells (e.g., T cells, B-cells, TILs (e.g., tumor infiltrating T-cells), including BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6, particularly within the context of certain diseases, including, e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer. As such, a step of detecting (or having detected) one or more somatic mutation in a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6) expressed by one or more subsets of immune cells (e.g., T cells, B-cells, etc.) is useful in methods of treating e.g., proinflammatory diseases (e.g., autoimmune diseases); and agents capable of inhibiting expression of one or more of these genes (BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) (e.g., RNA agents described herein) are useful in e.g., methods of treating e.g., proinflammatory diseases (e.g., autoimmune diseases). As such, the current disclosure provides, inter alia, methods of treating diseases associated with expression of one or more of said genes (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer) comprising detecting one or more somatic mutation in a target gene expressed by one or more subsets of immune cells (e.g., T cells, B-cclls, etc.); agents (e.g., RNA agents (e.g., RNAi agents, dsRNAi agents comprising a sense strand and an antisense strand, ssRNA agents)) capable of inhibiting gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6) expression (e.g., in a cell, e.g., in a cell in a subject) and their use in, inter alia, pharmaceutical compositions, and methods of treating diseases (e.g., proinflammatory diseases (e.g., autoimmune diseases), cancer).
TABLE OF CONTENTS
4.1 Definitions
4.2 RNA Agents
4.2.1 RNAi Agents
4.2.1.1 Antisense Strand
4.2.1.1 (i) T argeting Region
4.2.1.1(ii) Overall Length
4.2.1.1(iii) Exemplary Antisense Strands
4.2.1.2 Sense Strand
4.2.1.2(i) Antisense Strand Complementarity
4.2.1.2(ii) Overall Length
4.2.1.2(iii) Exemplary Sense Strands
4.2.1.3 dsRNA Agents
4.2.1.3(i) Single & Multiple Nucleic Acid Molecules
4.2.1.3(ii) Length of Double Stranded Region
4.2.1.3(iii) Nucleotide Overhangs & Blunt Ends
4.2.1.3(iv) Exemplary Structural Combinations of Sense & Antisense Strands
4.2.1.3(v) Exemplary Antisense Strands & Sense Strands
4.2.1.3(vi) Exemplary dsRNA Agents
4.2.2 Antisense Oligonucleotide RNA Agents
4.2.2.1 Targeting Region
4.2.2.2 Overall Length
4.2.2.3 Exemplary Antisense Oligonucleotides
4.3 Modified RNA Agents
4.3.1 Nature of Nucleotide Modifications 4.13 Exemplary Diseases
4.14 Exemplary Therapeutic Agents
4.15 Methods of Use
4.15.1 Methods of Treating, Ameliorating, or Preventing a Disease in a Subject
4.15.2 Methods of Diagnosing and/or Prognosticating a Disease in a Subject
4.15.3 Methods of Screening, Identifying, and Selecting a Subject for Treatment with a Therapeutic Agent
4.15.4 In Vitro Methods of Screening Samples for Somatic Gene Mutations
4.15.5 Methods of Delivery
4.15.6 Methods of Reducing or Inhibiting Gene Expression
4.16 Kits
4.1 Definitions
[0093] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.
[0094] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed.
[0095] In this application, the use of the singular includes the plural unless specifically stated otherwise. For example, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.
[0096] It is understood that wherever aspects are described herein with the language “comprising,” otherwise analogous aspects described in terms of “consisting of’ and “consisting essentially of’ are also provided and vice versa.
[0097] The term “and/or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and/or” as used in a phrase such as “A and/or B” herein is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Likewise, the term “and/or” as used in a phrase such as “A, B, and/or C” is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).
[0098] As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated.
[0099] The term “about” refers to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. When particular values or compositions are provided in the application and claims, unless otherwise stated, the meaning of “about” should be assumed to be within an acceptable error range for that particular' value or composition.
[00100] As used herein, the term “administering” refers to the physical introduction of an agent (e.g., an RNA agent (e.g., described herein)) e.g., a therapeutic agent (or a precursor of the therapeutic agent that is metabolized or altered within the body of the subject to produce the therapeutic agent in vivo) to a subject, using any of the various methods and delivery systems known to those skilled in the art. Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.
[00101] As used herein, the terms “agent” and “moiety” are used interchangeably herein and are used generically to describe any macro or micro molecule. Exemplary agents include, but are not limited polynucleotides (e.g., DNA, RNA (e.g., RNA)), proteins, peptides, small molecules, carbohydrates, lipids, synthetic polymers (e.g., polymers of PEG).
[00102] As used herein, the term “antisense strand” refers to an RNA molecule (e.g., in some embodiments, part of an RNAi agent (e.g., described herein), part of a dsRNA agent (e.g., described herein)) that comprises a region of complementarity comprising a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to a target nucleic acid sequence (e.g., a target mRNA (e.g., a BTG1 mRNA), a portion of a target mRNA (e.g., a BTG1 mRNA)). In some embodiments, the antisense strand is part of a dsRNA molecule that further comprises a sense strand. In some embodiments, the antisense strand or antisense oligonucleotide is a ssRNA agent. [00103] As used herein, the term “bicyclic sugar” refers to a modified sugar (e.g., ribose) moiety comprising two rings, wherein the second ring is formed via a bridge connecting two of the atoms in the first ring thereby forming a bicyclic structure. In some embodiments, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In some embodiments, the furanosyl sugar moiety is a ribosyl moiety.
[00104] As used herein, the term “bicyclic nucleoside” (“BNA”) is a nucleoside comprising a bicyclic sugar.
[00105] As used herein, the term “blunt end” refers to a dsRNA molecule that does not contain any unpaired nucleotides at the end (e.g., 3' terminus, 5' terminus) of the dsRNA molecule (z.e., no nucleotide overhang(s)). The dsRNA molecule can have, for example, a blunt end at the 3' end, 5' end, or both the 3' and 5' end of the molecule.
[00106] As used herein, the term “complementary” in reference to a first nucleotide sequence (e.g., a sense strand or a target mRNA) in relation to a second nucleotide sequence (e.g., an antisense strand), refers to the ability of a nucleic acid molecule comprising the first nucleotide sequence to hybridize to a nucleic acid molecule comprising the second nucleotide sequence and form a double stranded region (through base pair hydrogen bonds) under suitable in vivo or vitro conditions (e.g., under certain standard conditions, under mammalian (e.g., human) physiological conditions). A person of ordinary skill in the art would be able to select the set of conditions most appropriate for a hybridization test. Complementary sequences include, e.g., Watson-Crick base pairs. For example, complementary nucleobase pairs include adenine (A) and thymine (T); adenine (A) and uracil (U); and cytosine (C) and guanine (G). Complementary nucleobase pairs include natural and modified nucleotides, and nucleotide mimics, at least to the extent that the above hybridization requirements are fulfilled. As such, determinations of complementarity (as described herein) are independent of nucleotide chemical modifications (e.g., as described herein). For example, (C) and 5-methyl cytosine (mC) are both complementary to (G).
[00107] As used herein, the term “conjugation” refers to chemical conjugation of an RNA agent (e.g., a nucleic acid molecule) with another agent (e.g., carbohydrate, small molecule, polypeptide, polynucleotide, lipid, synthetic polymer (e.g., polymers of polyethylene glycol (PEG)), etc.). The other agent can be directly connected to the RNA agent (e.g., nucleic acid molecule) or indirectly connected through a linker, e.g., as described herein. Chemical conjugation methods are well known in the art, as are commercially available conjugation reagents and kits, with detailed instructions for their use readily available from the commercial suppliers.
[00108] As used herein, the term “differing by no more than X nucleotides” and the like (e.g., differing by not more than X nucleotides” in reference to a nucleotide sequence means that the nucleotide sequence comprises no more than X (wherein X is a specified number e.g., 3, 2, 1, 0)) nucleotide variations (as defined herein) relative to a reference sequence. For example, the phrase “wherein the nucleotide sequence of the antisense strand differs by no more than 3 nucleotides from the nucleotide sequence of SEQ ID NO: X” means that the nucleotide sequence comprises no more than 3 nucleotide variations relative to the nucleotide sequence set forth in the cited SEQ ID NO: X.
[00109] As used herein, the term “disease” refers to any abnormal condition that impairs physiological function. The term is used broadly to encompass any disorder, illness, abnormality, pathology, sickness, condition, or syndrome in which physiological function is impaired, irrespective of the nature of the etiology. The term disease includes infection (e.g., a viral, bacterial, fungal, protozoal infection).
[00110] As used herein, the term “double stranded RNA agent” or “dsRNA agent” refers to a complex of two RNA molecules comprising a double stranded region comprising two anti-parallel and at least partially (e.g., substantially, fully) complementary nucleic acid sequences that form the double stranded region. For example, in some embodiments, the dsRNA agent comprises a sense strand and an antisense strand.
[00111] As used herein, the term “BTG Anti-Proliferation Factor 1” or “BTG1” refers to the BTG/transducer of Erb family (Tob) member that functions, e.g., in the regulation of cell cycle progression, cell proliferation, and apoptosis. The term BTG1 includes human BTG1 (hBTGl). The mRNA sequence of reference hBTGl gene is set forth in SEQ ID NO: 1 (NCBI Ref.: NM_001731.3). The amino acid sequence of a reference hBTGl protein is set forth in SEQ ID NO: 3 (NCBI Ref.: NP_001722.1). The term BTGI includes naturally occurring variants of BTG1. BTG1 gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
[00112] As used herein, the term “ cyclin-dependent kinase inhibitor IB” or “CDKN1B” refers to the cyclin-dependent kinase inhibitor that belongs to the Cip/Kip family of cyclin dependent kinase inhibitor proteins. The term CDKN1B includes human CDKN1B (hCDKNIB). The mRNA sequence of reference hCDKNIB gene is set forth in SEQ ID NO: 4 (NCBI Ref.: NM_004064.5). The amino acid sequence of a reference hCDKNIB protein is set forth in SEQ ID NO: 6 (NCBI Ref.: NP_004055.1). The term CDKN1B includes naturally occurring variants of CDKN1B. CDKN1B gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
[00113] As used herein, the term “dual specificity phosphatase 2” or “DUSP2” refers to the dual specificity protein phosphatase that functions, e.g., in the inactivation of ERK1/2 and p38 MAPK. The term DUSP2 includes human DUSP2 (hDUSP2). The mRNA sequence of reference hDUSP2 gene is set forth in SEQ ID NO: 7 (NCBI Ref.: NM_004418.4). The amino acid sequence of a reference hDUSP2 protein is set forth in SEQ ID NO: 9 (NCBI Ref.: NP_004409.1). The term DUSP2 includes naturally occurring variants of DUSP2. DUSP2 gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
[00114] As used herein, the term “ immunoglobulin lambda like polypeptide 5” or “IGLL5” refers to the gene (and encoded protein) located within the immunoglobulin lambda locus, wherein the first exon of the IGLL5 gene is unrelated to immunoglobulin variable genes; while the second and third exons are the immunoglobulin lambda joining 1 and the immunoglobulin lambda constant 1 gene segments. The term IGLL5 includes human IGLL5 (hIGLL5). The mRNA sequence of reference hIGLL5 gene (isoform 1) is set forth in SEQ ID NO: 10 (NCBI Ref.: NM_001178126.2). The amino acid sequence of a reference hIGLL5 protein (isoform 1) is set forth in SEQ ID NO: 12 (NCBI Ref.: NP_001171597.1). The mRNA sequence of reference hIGLL5 gene (isoform 2) is set forth in SEQ ID NO: 4 (NCBI Ref.: NM_001256296.2). The amino acid sequence of a reference hIGLL5 protein (isoform 2) is set forth in SEQ ID NO: 15 (NCBI Ref.: NP_001243225.1). The term IGLL5 includes naturally occurring variants of IGLL5. IGLL5 gene and mRNA sequences of e.g., human, mouse, rat, non-human primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site. [00115] As used herein, the term “kelch like family member 6” or “KLHL6” refers to the KLHL family member that functions, e.g., in B-lymphocytc antigen receptor signaling and germinal- center B-cell maturation. The mRNA sequence of reference hKLHL6 gene is set forth in SEQ ID NO: 16 (NCBI Ref.: NM_ 130446.4). The amino acid sequence of a reference hKLHL6 protein is set forth in SEQ ID NO: 18 (NCBI Ref.: NP_569713.2). The term KLHL6 includes naturally occurring variants of KLHL6. KLHL6 gene and mRNA sequences of e.g., human, mouse, rat, nonhuman primate (e.g., rhesus macaque, Macaca fascicularis (cynomolgus monkey)), are readily available through publicly available databases, including, e.g., GenBank, UniProt, OMIM, and the Macaca genome project web site.
[00116] The terms “DNA” and “polydeoxyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple deoxyribonucleotides that are polymerized via phosphodiester bonds. Deoxyribonucleotides are nucleotides in which the sugar is deoxyribose.
[00117] As used herein, the term “fully complementary” means that in a hybridized pair of a first nucleic acid molecule and a second nucleic acid molecule, 100% (all), of the bases in a contiguous sequence of the first nucleic acid molecule will hybridize with the same number of bases in a contiguous sequence of the second nucleic acid molecule. The contiguous sequence may comprise all or a part of the first and/or second nucleic acid molecule.
[00118] As used herein, the term “heterologous,” when used to describe a first element in reference to a second element means that the first element and second element do not exist in nature disposed as described. For example, a nucleic acid molecule comprising a “heterologous moiety” means a nucleic acid molecule that is joined to a moiety (e.g., carbohydrate, small molecule, polypeptide, polynucleotide, lipid, synthetic polymer (e.g., polymers of PEG), etc.) that is not joined to the nucleic acid molecule in nature.
[00119] As used herein, the term “isolated” with reference to a polypeptide, protein, or polynucleotide refers to a polypeptide, protein, or polynucleotide that is substantially free of other cellular components with which it is associated in the natural state.
[00120] As used herein, the term “nucleotide variation,” “variant nucleotide,” or use of the term “variation” and the like in reference to a nucleotide or nucleic acid sequence refers to a nucleic acid molecule that comprises at least one substitution, addition, or deletion of one or more nucleotide compared to a reference nucleic acid molecule. As used herein, the term “variant” or “variation” with reference to a peptide or protein refers to a peptide or protein that comprises at least one substitution, addition, or deletion of an amino acid residue compared to a reference peptide or protein.
[00121] As used herein, the term “modified RNA agent” and the like refers to any RNA agent (or any component thereof (e.g., any nucleic acid molecule thereof)) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) described herein that comprises one or more modified nucleotide (as defined herein).
[00122] As used herein, the term “modified nucleotide,” “nucleotide modification,” or use of the term “modification” and the like in reference to a nucleotide or nucleic acid sequence refers to a nucleotide comprising a chemical modification, e.g., a modified sugar moiety, a modified nucleobase, and/or a modified internucleoside linkage, or any combination thereof. Exemplary modifications are provided herein, see, e.g., §§ 4.3, 4.3.1. In certain embodiments of the instant disclosure, inclusion of a deoxynucleotide - which is acknowledged as a naturally occurring form of nucleotide - if present within an RNA agent (e.g., RNAi agent) or component thereof (e.g., described herein, e.g., a sense strand, an antisense strand, a dsRNA agent) is considered to constitute a modified nucleotide.
[00123] As used herein, the term “nucleotide overhang” refers to at least one unpaired nucleotide that extends from the double stranded region of a nucleic acid molecule (e.g., a dsRNA molecule (e.g., a dsRNA molecule described herein)). For example, when a 3'-end of one strand of a dsRNA extends beyond the 5'-end of the other strand, or vice versa, there is a nucleotide overhang.
[00124] As used herein, the term, “non-complementary nucleotide mismatch” refers to a nucleotide within a region of complementarity (as described herein) that is not complementary to the corresponding nucleotide in the target nucleic acid molecule.
[00125] As used herein, the term “obtaining a sample” refers to the acquisition of a sample. The term includes the direct acquisition from a subject and the indirect acquisition through one or more third parties wherein one of the third parties directly acquired the sample from the subject.
[00126] As used herein, the term “operably connected” refers to the linkage of two moieties in a functional relationship. For example, a polypeptide is operably connected to another polypeptide when they are linked (either directly or indirectly via a peptide linker) in frame such that both polypeptides are functional (e.g., a fusion protein described herein). Or for example, a transcription regulatory polynucleotide e.g., a promoter, enhancer, or other expression control element is operably linked to a polynucleotide that encodes a protein if it affects the transcription of the polynucleotide that encodes the protein. The term “operably connected” can also refer to the conjugation of a moiety to e.g., a polynucleotide or polypeptide (e.g., the conjugation of a PEG polymer to a protein).
[00127] As used herein, “partially complementary” means that in a hybridized pair of a first nucleic acid molecule and a second nucleic acid molecule, at least 70%, but not all, of the bases in a contiguous sequence of the first nucleic acid molecule will hybridize with the same number of bases in a contiguous sequence of the second nucleic acid molecule. The contiguous sequence may comprise all or a part of a first or second nucleic acid molecule.
[00128] The determination of “percent identity” between two sequences (e.g., protein (amino acid sequences) or polynucleotide (nucleic acid sequences)) can be accomplished using a mathematical algorithm. Determinations of identity (as described herein) are independent of nucleotide chemical modifications (e.g., as described herein). For example, (mC) is identical to (C) for the purposes of determining percent identity. A specific, non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87: 2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90: 5873-5877, the entire contents of each of which are incorporated herein by reference in their entirety for all purposes. Such an algorithm is incorporated into the NBEAST and XBEAST programs of Altschul SF et al., (1990) J Mol Biol 215: 403, which is incorporated herein by reference in its entirety for all purposes. BEAST nucleotide searches can be performed with the NBEAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecule described herein. BEAST protein searches can be performed with the XBEAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule described herein. To obtain gapped alignments for comparison purposes, Gapped BEAST can be utilized as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, which is incorporated herein by reference in its entirety. Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17, which is incorporated herein by reference in its entirety. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing amino acid sequences, a PAM 120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used. The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. In calculating percent identity, typically only exact matches are counted.
[00129] As used herein, the term “pharmaceutical composition” means a composition that is suitable for administration to an animal, e.g., a human subject, and comprises a therapeutic agent and a pharmaceutically acceptable carrier or diluent.
[00130] As used herein, the term “pharmaceutically acceptable carrier or diluent” means a substance intended for use in contact with the tissues of human beings and/or non-human animals, and without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable therapeutic benefit/risk ratio.
[00131] The terms “nucleic acid molecule,” “polynucleotide,” and “oligonucleotide” are used interchangeably herein and refer to a polymer of DNA or RNA. The nucleic acid molecule can be single- stranded or double-stranded; contain natural, non-natural, or modified nucleotides; and contain a natural, non-natural, or modified intemucleoside linkage, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule. Nucleic acid molecules include, but are not limited to, all nucleic acid molecules which are obtained by any means available in the art, including, without limitation, recombinant means, e.g., the cloning of nucleic acid molecules from a recombinant library or a cell genome, using ordinary cloning technology and polymerase chain reaction, and the like, and by synthetic means. The skilled artisan will appreciate that, except where otherwise noted, nucleic acid sequences set forth in the instant application will recite thymidine (T) in a representative DNA sequence but where the sequence represents RNA (e.g., mRNA), the thymidines (Ts) would be substituted for uracils (Us). Thus, any of the RNA polynucleotides encoded by a DNA identified by a particular sequence identification number may also comprise the corresponding RNA (e.g., mRNA) sequence encoded by the DNA, where each thymidine (T) of the DNA sequence is substituted with uracil (U). [00132] As used herein, the term “plurality” means 2 or more (e.g., 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 9 or more, or 10 or more).
[00133] As used herein, the terms “protein” and “polypeptide” refers to a polymer of at least 2 (e.g., at least 5) amino acids linked by a peptide bond. The term “polypeptide” does not denote a specific length of the polymer chain of amino acids. It is common in the art to refer to shorter polymers of amino acids e.g., approximately 2-50 amino acids) as peptides; and to refer to longer polymers of amino acids (e.g., approximately over 50 amino acids) as polypeptides. However, the terms “peptide” and “polypeptide” and “protein” are used interchangeably herein. In some embodiments, the protein is folded into its three-dimensional structure. Where proteins are contemplated herein, it should be understood that proteins folded into their three-dimensional structure are also provided herein as well as polypeptides in the primary structure. Proteins can include more than one polypeptide (e.g., customarily referred to as the quaternary structure).
[00134] As used herein, the term “prognostication” and the like refers to the process of estimating/predicting the likely course and/or outcome of a disease in a subject, including, e.g., the chance that a subject has of recovering from the disease.
[00135] As used herein, the term “region of complementarity” refers to a portion of a first nucleic acid molecule comprising a nucleotide sequence that is at least partially complementary to the nucleotide sequence of at least a portion of a second nucleic acid molecule.
[00136] The terms “RNA” and “polyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple ribonucleotides that are polymerized via phosphodiester bonds. Ribonucleotides are nucleotides in which the sugar’ is ribose. RNA may contain modified nucleotides; and contain natural, non-natural, or modified internucleoside linkages, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule.
[00137] As used herein, the term “RNAi agent” refers to an RNA agent that contains one or more RNA molecules which can mediate the targeted cleavage of an RNA molecule (e.g., an mRNA molecule) via an RNA-induced silencing complex (RISC) pathway. The RNAi agent, is thereby capable of e.g., modulating, e.g., inhibiting, the expression of a target gene (e.g., BTG1) in a cell, e.g., a cell within a subject, such as a mammalian subject. In some embodiments, the RNAi agent is a dsRNA agent comprising a sense strand and an antisense strand that form a double stranded region. In some embodiments, the sense strand and the antisense strand each independently comprise or consist of from about 19-23 nucleotides.
[00138] As used herein, the term “sample” encompasses a variety of biological specimens obtained from a subject. Exemplary sample types include, e.g., blood, red blood cells, and other liquid samples of biological origin (including, but not limited to, whole-blood, red blood cells e.g., isolated from whole blood), peripheral blood mononuclear' cells (PBMCs), serum, plasma, urine, saliva, amniotic fluid, stool, synovial fluid, etc.), nasopharyngeal swabs, solid tissue samples such as biopsies (or cells derived therefrom and the progeny thereof), tissue cultures (or cells derived therefrom and the progeny thereof), and cell cultures (or cells derived therefrom and the progeny thereof). The term also includes samples that have been manipulated in any way after their procurement from a subject, such as by centrifugation, filtration, washing, precipitation, dialysis, chromatography, lysis, treatment with reagents, enriched for certain cell populations, refrigeration, freezing, staining, etc.
[00139] As used herein, the term “sense strand” refers to an RNA molecule (e.g., part of an RNAi agent (e.g., described herein), part of a dsRNA agent (e.g., described herein)) that comprises a region that is at least partially (e.g., substantially, fully) complementary to a region of the antisense strand (as defined herein). The sense strand is often referred to as such with reference to the orientation of the sequence of the sense strand being the same with respect to a target RNA (e.g., mRNA sequence).
[00140] As used herein, the term “somatic mutation” refers to one or more nucleotide alteration (e.g., variation, modification (e.g., variation)) in a genomic DNA sequence (e.g., of a gene) that is acquired during the lifetime of a subject. It is understood in the art that somatic mutations are not present in the germline DNA of an individual and are therefore not inherited from a parent like germline polymorphisms. Somatic mutations may occur spontaneously due to e.g., the infidelity of DNA replication occurring at each cell division creating substitutions, deletions, or additions of nucleotides into the DNA of a cell. A somatic mutation may also be caused by environmental factors such as e.g., ultraviolet radiation, chemical exposure, or virial infections.
[00141] As used herein, the term “subject” includes any animal, such as a human or other animal. In some embodiments, the subject is a vertebrate animal (e.g., mammal, bird, fish, reptile, or amphibian). In some embodiments, the subject is a human. In some embodiments, the method subject is a non-human mammal. In some embodiments, the subject is a non-human mammal is such as a non-human primate (e.g., monkeys, apes), ungulate (e.g., cattle, buffalo, sheep, goat, pig, camel, llama, alpaca, deer, horses, donkeys), carnivore (e.g., dog, cat), rodent (e.g., rat, mouse), or lagomorph (e.g., rabbit). In some embodiments, the subject is a bird, such as a member of the avian taxa Galliformes (e.g., chickens, turkeys, pheasants, quail), Anseriformes (e.g., ducks, geese), Paleaognathae (e.g., ostriches, emus), Columbiformes (e.g., pigeons, doves), or Psittaciformes (e.g., parrots).
[00142] As used herein, “substantially complementary” means that in a hybridized pair of a first nucleic acid molecule and a second nucleic acid molecule, at least 85%, but not all, of the bases in a contiguous sequence of the first nucleic acid molecule will hybridize with the same number of bases in a contiguous sequence of the second nucleic acid molecule. The contiguous sequence may comprise all or a pail of a first or second nucleic acid molecule.
[00143] In some embodiments, the term “substantially all” means at least 95%, 96%, 97%, 98% or 99%, e.g., of the subject of said sentence, but excluding 100% (i.e., all).
[00144] As used herein, the term “target nucleic acid sequence” refers to a contiguous portion of the nucleotide sequence of a nucleic acid sequence (e.g., an mRNA molecule formed during the transcription of a target gene (e.g., BTG1)). In some embodiments, the target nucleic acid sequence is an mRNA molecule formed during the transcription of a target gene (e.g., BTG1)). In some embodiments, the target nucleic acid molecule comprises an mRNA that is a product of RNA processing of a primary transcription product. In some embodiments, the target portion of the sequence (e.g., mRNA) will be at least long enough to serve as a substrate for RNAi-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a BTG1 gene. In one embodiment, the target sequence is within the protein coding region of BTG1.
[00145] As used herein, the term “therapeutically effective amount” of a therapeutic agent refers to any amount of the therapeutic agent that, when used alone or in combination with another therapeutic agent, improves a disease condition, e.g., protects a subject against the onset of a disease; improves a symptom of disease, e.g., decreases severity of disease symptoms, decreases frequency or duration of disease symptoms, increases disease symptom- free periods; prevents or reduces impairment or disability due to the disease; or promotes disease (or infection) regression. The ability of a therapeutic agent to improve a disease condition can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the RNA agent in in vitro assays.
[00146] As used herein, the terms “treat,” treating,” “treatment,” and the like refer to reducing or ameliorating a disease and/or symptom(s) associated therewith or obtaining a desired pharmacologic and/or physiologic effect. It will be appreciated that, although not precluded, treating a disease does not require that the disease, or symptom(s) associated therewith be completely eliminated. In some embodiments, the effect is therapeutic, i.e., without limitation, the effect partially or completely reduces, diminishes, abrogates, abates, alleviates, decreases the intensity of, or cures a disease and/or adverse symptom attributable to the disease. In some embodiments, the effect is preventative, i.e., the effect protects or prevents an occurrence or reoccurrence of a disease. To this end, the presently disclosed methods comprise administering a therapeutically effective amount of a compositions as described herein.
4.2 RNA Agents
[00147] Provided herein are inter alia, agents (e.g., RNA agents), useful in, inter alia, inhibiting expression of one or more target gene (e.g., expressed by one or more subsets of immune cells, e.g., T cells) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of mRNA encoded by the target gene).
[00148] Target genes include, BTG1 (e.g., human BTG1 (hBTGl)); CDKN1B (e.g., human CDKN1B (hCDKNIB)); DUSP2 (e.g., human DUSP2 (hDUSP2)); IGLL5 (e.g., human IGLL5 (hIGLL5)); or KLHL6 (e.g., human KLHL6 (hKLHL6)).
[00149] As such provided herein are, inter alia, RNA agents (e.g., RNAi agents, dsRNA agents, ssRNA agents, siRNA agents, antisense oligonucleotides), useful in, inter alia, inhibiting expression of: BTG1 (BTG1) (e.g., human BTG1 (hBTGl)) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of BTG1 (e.g., hBTGl) mRNA); CDKN1B (e.g., hCDKNIB) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of CDKN1B (e.g., hCDKNIB) mRNA); DUSP2 (e.g., hDUSP2) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of DUSP2 (e.g., hDUSP2) mRNA); IGLL5 (e.g., hIGLL5) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of IGLL5 (e.g., hIGLL5) mRNA); or KLHL6 (e.g., hKLHL6) (e.g., within a cell, e.g., within a cell in a subject, e.g., a mammalian subject, e.g., a human subject) (e.g., through the degradation of KLHL6 (e.g., hKLHL6) mRNA).
[00150] BTG1 is a member of the BTG/ Tob family, generally regarded as an anti-proliferative gene family. The BTG/Tob family consists of six protein members; BTG1, BTG2/Tis21/PC3, BTG3/ANA, BTG4/PC3B, TOB1/TOB, and TOB2. BTG1 functions in a wide range of cellular activities including, for example, proliferation, apoptosis, and cell growth. BTG1 expression is generally highest in the G0/G1 phases of the cell cycle and downregulated as cells progress through Gl. BTG1 is expressed both in the nucleus and cytoplasm, and function influenced at least in part by the intracellular localization. In the nuclease BTG1 is known to interact with several nuclear receptors and can function as a coactivator of cell differentiation. See, e.g., Kim, Sang Hyeon et al. “Emerging role of anti-proliferative protein BTG1 and BTG2.” BMB reports vol. 55,8 (2022): 380-388. doi:10.5483/BMBRep.2022.55.8.092. The mRNA sequence of a reference hBTGl gene is set forth in SEQ ID NO: 1. The reverse complement sequence of the hBTGl mRNA is set forth in SEQ ID NO: 2. The amino acid sequence of the hBTGl protein encoded by the hBTGl reference gene is set forth in SEQ ID NO: 3.
[00151] CDKN1B is a cyclin-dependent kinase inhibitor that belongs to the Cip/Kip family of cyclin dependent kinase inhibitor proteins. CDKN1B functions to, e.g., bind to and prevent the activation of cyclin E-CDK2 or cyclin D-CDK4 complexes, and thus control the cell cycle progression at the Gl phase. CDKN1B degradation, which is triggered by, e.g., its CDK dependent phosphorylation and subsequent ubiquitination by SCF complexes (Skp, Cullin, F-box containing complex), is generally needed for the cellular transition from quiescence to a proliferative state. The mRNA sequence of a reference hCDKNIB gene is set forth in SEQ ID NO: 4. The reverse complement sequence of the hCDKNIB mRNA is set forth in SEQ ID NO: 5. The amino acid sequence of the hCDKNIB protein encoded by the hCDKNIB reference gene is set forth in SEQ ID NO: 6.
[00152] DUSP2 is a member of the dual specificity protein phosphatase subfamily. These phosphatases inactivate their target kinases by dephosphorylating both phosphoserine/threonine and pho sphotyro sine residues. Members of the DUSP family exhibit distinct substrate specificities for various MAP kinases, different tissue distribution, different subcellular localization, and different modes of inducibility of their expression by extracellular stimuli. DUSP2, for example, inactivates ERK1/2 and p38 MAPK. DUSP2 is expressed in, e.g., hematopoietic tissues, and is localized in the nucleus. The mRNA sequence of a reference hDUSP2 gene is set forth in SEQ ID NO: 7. The reverse complement sequence of the hDUSP2 mRNA is set forth in SEQ ID NO: 8. The amino acid sequence of the hDUSP2 protein encoded by the hDUSP2 reference gene is set forth in SEQ ID NO: 9.
[00153] The IGLL5 gene is located within the immunoglobulin lambda locus but it does not require somatic rearrangement for expression. The first exon of the IGLL5 gene is unrelated to immunoglobulin variable genes; while the second and third exons are the immunoglobulin lambda joining 1 and the immunoglobulin lambda constant 1 gene segments. It is known that alternative splicing results in multiple transcript variants. The mRNA sequence of a reference hIGLL5 gene (isoform 1) is set forth in SEQ ID NO: 10. The reverse complement sequence of the hIGLL5 mRNA (isoform 1) is set forth in SEQ ID NO: 11. The amino acid sequence of the hIGLL5 protein encoded by the hIGLL5 (isoform 1) reference gene is set forth in SEQ ID NO: 12. The mRNA sequence of a reference hIGLL5 gene (isoform 2) is set forth in SEQ ID NO: 13. The reverse complement sequence of the hIGLLS mRNA (isoform 2) is set forth in SEQ ID NO: 14. The amino acid sequence of the hIGLL5 protein encoded by the hIGLL5 (isoform 2) reference gene is set forth in SEQ ID NO: 15.
[00154] KLHL6 is a member of the kelch-like (KLHL) family of proteins. Proteins of the KLHL family generally contain a BTB/POZ domain, a BACK domain, and five to six Kelch motifs. The KLHL6 protein contains an N-terminal broad-complex, tramtrack and brie a brae (BTB) domain that facilitates protein binding and dimerization, a BTB domain, C-terminal kelch (BACK) domain, and six C-terminal kelch repeat domains. KLHL6 functions, e.g., in B-lymphocyte antigen receptor signaling and germinal-center B-cell maturation. KLHL6 is expressed by lymphoid tissue, including, e.g., by germinal center B-cells. The mRNA sequence of a reference hKLHL6 gene is set forth in SEQ ID NO: 16. The reverse complement sequence of the hKLHL6 mRNA is set forth in SEQ ID NO: 17. The amino acid sequence of the hKLHL6 protein encoded by the hKLHL6 reference gene is set forth in SEQ ID NO: 18.
Table 1. The mRNA and Amino Acid Sequence of a Reference hBTGl, hCDKN 1 B, hDUSP2, hIGLL5, and hKLHL6,
[00155] In some embodiments, the RNA agent comprises one or more RNA molecule. In some embodiments, the RNA agent is an RNAi agent. In some embodiments, the RNAi agent is a dsRNA agent. In some embodiments, the RNA agent is an siRNA agent. In some embodiments, the RNA agent comprises a dsRNA agent. In some embodiments, the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand. In some embodiments, the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand that form a double stranded region. In some embodiments, the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand that hybridize to form a double stranded region. In some embodiments, the sense strand and the antisense strand are part of a single nucleic acid molecule (e.g., a single nucleic acid molecule comprising a hairpin loop). In some embodiments, the sense strand and the antisense strand are separate nucleic acid molecules. In some embodiments, the RNA agent comprises an antisense strand. In some embodiments, the RNA agent comprises a sense strand. In some embodiments, the RNA agent comprises one or more single stranded RNA (ssRNA) molecule. In some embodiments, the RNA agent consists of an antisense strand. In some embodiments, the RNA agent is an antisense oligonucleotide agent.
4.2.1 RNAi Agents
[00156] In some embodiments, the RNA agent is an RNAi agent. In some embodiments, the RNAi agent comprises an antisense strand and a sense strand. In some embodiments, the RNAi agent is a double stranded RNA (dsRNA) agent comprising a sense strand and an antisense strand. In some embodiments, the RNAi agent is an siRNA agent. 4.2.1.1 Antisense Strand
(i) Targeting Region
[00157] Antisense strands (e.g., described herein) comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is at least substantially complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is fully complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, 1GLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
[00158] In some embodiments, the nucleotide sequence of the region of complementarity is at least 70%, 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). For example, the nucleotide sequence of the region of complementarity may be at least 70% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 75% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 80% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 85% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 90% complementary to the nucleotide sequence of the target nucleic acid molecule e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 95% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is at least 95%, 96%, 97%, 98%, 99%, or 100% (e.g., in some embodiments, preferably at least 95%, more preferably at least 98%) complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
[00159] In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of one or more non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 5 (e.g., 4, 3, 2, 1, or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 5 (e.g., 4, 3, 2, 1 , or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 3 (e. ., 2, 1, or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 2 (e.g., 1 or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 1 (e.g., 0) non-complementary nucleotide mismatch relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises 0 non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity. In some embodiments, the region of complementarity comprises at least one but not more than 3 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non- complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity. In some embodiments, the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity. In some embodiments, the region of complementarity comprises at least one but not more than 3 non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity. Methods known in the art and described herein can be utilized to evaluate the effect of any non- complementary mismatches between an antisense strand and a target nucleic acid molecule on functional properties (e.g., inhibition of expression of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTGl , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA))).
[00160] In some embodiments, the region of complementarity comprises from about 15-30 nucleotides, e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30,
19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27,
20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides. In some embodiments, the region of complementarity comprises from about 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-
24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides. In some embodiments, the region of complementarity comprises from about 19-21 (e.g., 19-20) nucleotides. In some embodiments, the region of complementarity comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity comprises about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity comprises about 19 nucleotides. In some embodiments, the region of complementarity comprises about 20 nucleotides. In some embodiments, the region of complementarity comprises about 21 nucleotides. In some embodiments, the region of complementarity comprises about 22 nucleotides. In some embodiments, the region of complementarity comprises about 23 nucleotides. In some embodiments, the region of complementarity consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23,
24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity consists of about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity consists of about 19 nucleotides. In some embodiments, the region of complementarity consists of about 20 nucleotides. In some embodiments, the region of complementarity consists of about 21 nucleotides. In some embodiments, the region of complementarity consists of about 22 nucleotides. In some embodiments, the region of complementarity consists of about 23 nucleotides. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
[00161] In some embodiments, the target nucleic acid molecule is part (e.g., a contiguous portion) of a larger nucleic acid molecule. For example, in some embodiments, the target nucleic acid molecule is a portion (e.g., a contiguous portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA). In some embodiments, the target nucleic acid molecule is a contiguous nucleotide sequence of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) of sufficient length to allow it to be a substrate for cleavage directed by an RNAi agent e.g., an RNAi agent described herein, e.g., a dsRNA agent (e.g., described herein)) (i.e., cleavage through a RISC pathway).
[00162] In some embodiments, the target nucleic acid molecule is a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA). In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA). In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) formed in the expression of a target gene (e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene) (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 gene).
[00163] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a BTG1 (e.g., hBTGl) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a BTG1 (e.g., hBTGl) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 1 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a BTG1 (e.g., hBTGl) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 3 (or a variant or fragment thereof).
[00164] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a CDKNIB (e.g., hCDKNIB) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a CDKNIB (e.g., hCDKNIB) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 4 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a CDKN1B e.g., hCDKNIB) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 6 (or a variant or fragment thereof).
[00165] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a DUSP2 (e.g., hDUSP2) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a DUSP2 (e.g., hDUSP2) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 7 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a DUSP2 (e.g., hDUSP2) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 9 (or a variant or fragment thereof).
[00166] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a IGLL5 (e.g., hIGLL5) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a IGLL5 (e.g., hIGLL5) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 10 (or a valiant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 12 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 13 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 15 (or a variant or fragment thereof).
[00167] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a KLHL6 (e.g., hKLHL6) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a KLHL6 (e.g., hKLHL6) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 16 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a KLHL6 (e.g., hKLHL6) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 18 (or a variant or fragment thereof).
[00168] In some embodiments, the target nucleic acid molecule comprises from about 19-30 nucleotides, e.g., 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20- 29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26,
21-25, 21-24, 21-23, 21-22, 22-30, 22-29, 22-28, 22-27, 22-26, 22-25, 22-24, 22-23, 23-30, 23-29,
23-28, 23-27, 23-26, 23-27, 23-26, 23-25, or 23-24 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 19-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 19-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 19, 18, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 19 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 20 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 21 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 23 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 19, 18, 20,
21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 19 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 20 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 21 nucleotides. Tn some embodiments, the target nucleic acid molecule consists of about 23 nucleotides.
[00169] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
(ii) Overall Length
[00170] In some embodiments, the antisense strand comprises from about 15-30 nucleotides (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17,
18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28,
19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25,
20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides). In some embodiments, the antisense strand comprises from about 18-25 nucleotides (e.g., 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20- 24,20-23, 20-22, 20-21 , 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides). In some embodiments, the antisense strand comprises from about 19-25 nucleotide (e.g., 19-20, 19-21, 19-22, 19-23, 19-24, 19-25, 20-21, 20-22, 20-23, 20-24, 20-25, 21-22, 21-23,
21-24, 21-25, 22-23, 22-24, 22-25, 23-24, 23-25, 24-25 nucleotides). In some embodiments, the antisense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-3020-30, 21-30, 22-30, 23- 30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides. In some embodiments, the antisense strand comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense strand comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense strand comprises about 21 nucleotides. In some embodiments, the antisense strand comprises about 23 nucleotides. In some embodiments, the antisense strand consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense strand consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense strand consists of about 21 nucleotides. In some embodiments, the antisense strand consists of about 23 nucleotides.
[00171] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
(iii) Exemplary Antisense Strands [00172] In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00173] In some embodiments, the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides. [00174] In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00175] In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32,
33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of a portion of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
[00176] In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,
34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 , 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00177] In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00178] In some embodiments, the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 19 contiguous nucleotides.
[00179] In some embodiments, the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 20 contiguous nucleotides.
[00180] In some embodiments, the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 21 contiguous nucleotides.
[00181] In some embodiments, the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 22 contiguous nucleotides.
[00182] In some embodiments, the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand comprises 23 contiguous nucleotides.
[00183] In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00184] In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00185] In some embodiments, the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 19 contiguous nucleotides.
[00186] In some embodiments, the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 20 contiguous nucleotides.
[00187] In some embodiments, the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 21 contiguous nucleotides.
[00188] In some embodiments, the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 22 contiguous nucleotides.
[00189] In some embodiments, the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense strand consists of 23 contiguous nucleotides.
[00190] In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00191] In some embodiments, the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 20 e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00192] In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00193] In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides. [00194] In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00195] In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00196] In some embodiments, the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 19 contiguous nucleotides.
[00197] In some embodiments, the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 20 contiguous nucleotides.
[00198] In some embodiments, the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 21 contiguous nucleotides.
[00199] In some embodiments, the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 22 contiguous nucleotides.
[00200] In some embodiments, the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand comprises 23 contiguous nucleotides.
[00201] In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 15, 16, 17, 18,
19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,
29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00202] In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27,
28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,
30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of
20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28,
29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00203] In some embodiments, the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11 , 14, or 17. In some embodiments, the antisense strand consists of 19 contiguous nucleotides differing by no more than 5 {e.g., 0, 1, 2, 3, 4, or 5) {e.g., by no more than 3 {e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 19 contiguous nucleotides.
[00204] In some embodiments, the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 {e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 20 contiguous nucleotides differing by no more than 5 {e.g., 0, 1, 2, 3, 4, or 5) {e.g., by no more than 3 {e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 20 contiguous nucleotides.
[00205] In some embodiments, the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 {e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 21 contiguous nucleotides differing by no more than 5 {e.g., 0, 1, 2, 3, 4, or 5) {e.g., by no more than 3 {e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 21 contiguous nucleotides.
[00206] In some embodiments, the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 {e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 22 contiguous nucleotides differing by no more than 5 {e.g., 0, 1, 2, 3, 4, or 5) {e.g., by no more than 3 {e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 22 contiguous nucleotides.
[00207] In some embodiments, the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 23 contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense strand consists of 23 contiguous nucleotides.
[00208] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
4.2.1.2 Sense Strand
(i) Antisense Strand Complementarity
[00209] As described above, sense strands (e.g., described herein) comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of at least a portion of an antisense strand. As such, pairs of sense and antisense strands can hybridize to form a double stranded region (e.g., under conditions in which the pairs will be used).
[00210] In some embodiments, the nucleotide sequence of the region of complementarity is at least substantially complementary to the nucleotide sequence of at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity is fully complementary to the nucleotide sequence of at least a portion of an antisense strand.
[00211] In some embodiments, the nucleotide sequence of the region of complementarity is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of at least a portion of an antisense strand. For example, the nucleotide sequence of the region of complementarity may be at least 70% complementary to the nucleotide sequence of at least a portion of an antisense strand. The nucleotide sequence of the region of complementarity may be at least 75% complementary to the nucleotide sequence of at least a portion of an antisense strand. The nucleotide sequence of the region of complementarity may be at least 80% complementary to the nucleotide sequence of at least a portion of an antisense strand. The nucleotide sequence of the region of complementarity may be at least 85% complementary to the nucleotide sequence of at least a portion of an antisense strand. The nucleotide sequence of the region of complementarity may be at least 90% complementary to the nucleotide sequence of at least a portion of an antisense strand. The nucleotide sequence of the region of complementarity may be at least 95% complementary to the nucleotide sequence of at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity is at least 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of at least a portion of an antisense strand.
[00212] In some embodiments, the nucleotide sequence of the region of complementarity comprises one or more non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 5 (e.g., 4, 3, 2, 1, or 0) non- complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 3 (e.g., 2, 1, or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 2 e.g., 1 or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 1 (e.g., 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the nucleotide sequence of the region of complementarity comprises 0 non- complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand. In some embodiments, the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand, wherein the one or more non- complementary nucleotide mismatch is within the last 5 (e.g., 4, 3, 2, or 1 ) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity. In some embodiments, the region of complementarity comprises at least one but not more than 3 (e.g., 1, 2, or 3) non-complementary nucleotide mismatches relative to the nucleotide sequence of the at least a portion of an antisense strand, wherein the one or more non-complementary nucleotide mismatch is within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity.
[00213] In some embodiments, the region of complementarity comprises from about 15-30 nucleotides, e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30,
19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27,
20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides. In some embodiments, the region of complementarity comprises from about 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20- 24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides. In some embodiments, the region of complementarity comprises from about 19-21 (e.g., 19-20) nucleotides. In some embodiments, the region of complementarity comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity comprises about 19, 20, or 21 nucleotides. In some embodiments, the region of complementarity comprises about 19 nucleotides. In some embodiments, the region of complementarity comprises about 20 nucleotides. In some embodiments, the region of complementarity comprises about 21 nucleotides. In some embodiments, the region of complementarity consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity consists of about 19, 20, or 21 nucleotides. In some embodiments, the region of complementarity consists of about 19 nucleotides. In some embodiments, the region of complementarity consists of about 20 nucleotides. In some embodiments, the region of complementarity consists of about 21 nucleotides.
[00214] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
(ii) Overall Length
[00215] In some embodiments, the sense strand comprises from about 15-30 nucleotides (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21 , 15-20, 15-19, 15-18, 15-17, 18- 30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28,
19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25,
20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides). In some embodiments, the sense strand comprises from about 18-25 nucleotides (e.g.,
18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-24,20-23,
20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides). In some embodiments, the sense strand comprises from about 19-25 nucleotide (e.g.,
19-20, 19-21, 19-22, 19-23, 19-24, 19-25, 20-21, 20-22, 20-23, 20-24, 20-25, 21-22, 21-23, 21-24,
21-25, 22-23, 22-24, 22-25, 23-24, 23-25, 24-25 nucleotides). In some embodiments, the sense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-30 20-30, 21-30, 22-30, 23-30, 24- 30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides. [00216] In some embodiments, the sense strand comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense strand comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense strand comprises about 19, 20, 21, 22, 23 nucleotides. In some embodiments, the sense strand comprises about 19, 20, 21 nucleotides. In some embodiments, the sense strand comprises about 20 nucleotides. In some embodiments, the sense strand comprises about 21 nucleotides. In some embodiments, the sense strand comprises about 21 nucleotides. In some embodiments, the sense strand comprises about 23 nucleotides.
[00217] In some embodiments, the sense strand consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense strand consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the sense strand consists of about 19, 20, 21, 22, 23 nucleotides. In some embodiments, the sense strand consists of about 19, 20, 21 nucleotides. In some embodiments, the sense strand consists of about 20 nucleotides. In some embodiments, the sense strand consists of about 21 nucleotides. In some embodiments, the sense strand consists of about 21 nucleotides. In some embodiments, the sense strand consists of about 23 nucleotides.
[00218] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
(iii) Exemplary Sense Strands [00219] In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00220] In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00221] In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00222] In some embodiments, the sense strand comprises at least 30 e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
[00223] In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a B TGI, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00224] In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of aportion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides. [00225] In some embodiments, the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 19 contiguous nucleotides.
[00226] In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 20 contiguous nucleotides.
[00227] In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 21 contiguous nucleotides.
[00228] In some embodiments, the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 22 contiguous nucleotides.
[00229] In some embodiments, the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand comprises 23 contiguous nucleotides.
[00230] In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00231] In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides. [00232] In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16.
[00233] In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 19 contiguous nucleotides.
[00234] In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 20 contiguous nucleotides.
[00235] In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 21 contiguous nucleotides.
[00236] In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 22 contiguous nucleotides.
[00237] In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the sense strand consists of 23 contiguous nucleotides.
[00238] In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00239] In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00240] In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27 , 28,
29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 25 (e.g., at least 26, 27, 28, 29,
30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00241] In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
[00242] In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35 contiguous nucleotides.
[00243] In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 {e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00244] In some embodiments, the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 19 contiguous nucleotides. [00245] In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 20 contiguous nucleotides. [00246] In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1 , 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 21 contiguous nucleotides. [00247] In some embodiments, the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 22 contiguous nucleotides. [00248] In some embodiments, the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises 23 contiguous nucleotides. [00249] In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1 , 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00250] In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,
32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,
33, 34, or 35 contiguous nucleotides.
[00251] In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 19 contiguous nucleotides. [00252] In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 20 contiguous nucleotides. [00253] In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 21 contiguous nucleotides. [00254] In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 22 contiguous nucleotides. [00255] In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS:
1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand consists of 23 contiguous nucleotides. [00256] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
4.2.1.3 dsRNA Agents
[00257] In some embodiments, the RNA agent (e.g., RNAi agent) comprises a dsRNA agent comprising an antisense strand (e.g., described herein, e.g., described in § 4.2.1.1) and a sense strand (e.g., described herein, e.g., described in § 4.2.1.2) that hybridize to form a double stranded region (e.g., under conditions in which the dsRNA will be used (e.g., under physiological (e.g., mammalian, e.g., human) conditions within a cell)). [00258] As described above, antisense strands (e.g., described herein) comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)); and the sense strands comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of at least a portion of an antisense strand.
(i) Single & Multiple Nucleic Acid Molecules
[00259] As described herein, and known in the art, the sense strand and the antisense strand can be part of a single larger nucleic acid molecule (connected as a single stranded nucleic acid molecule) or separate nucleic acid molecules (only connected through the double stranded region). In some embodiments, the sense strand and the antisense strand are separate nucleic acid molecules. In some embodiments, sense strand and the antisense strand are pail of a single larger nucleic acid molecule.
[00260] In embodiments wherein the sense and antisense strands are part of a single nucleic acid molecule, the nucleic acid molecule may comprise a hairpin loop between the antisense strand and the sense strand to allow for formation of the double stranded region. In some embodiments, the hairpin loop comprises at least 1 (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 23, 25 or more) unpaired nucleotides (non-complementary nucleotide mismatches). In some embodiments, the hairpin loop comprises at least one but less than 25, 23, 20, 10, 9, 8, 7, 6, 5, 4, 3, or 2 unpaired nucleotides (non-complementary nucleotide mismatches). In some embodiments, the hairpin loop comprises about 25, 23, 20, 9, 8, 7, 6, 5, 4, 3, or 1 unpaired nucleotide (non-complementary nucleotide mismatch). In some embodiments, the hairpin loop consists of about 25, 23, 20, 9, 8, 7, 6, 5, 4, 3, or 1 unpaired nucleotide (non-complementary nucleotide mismatch).
[00261] Without wishing to be bound by theory, in embodiments wherein the sense strand and the antisense strand are part of a single nucleic acid molecule, after introduction into a suitable cell (e.g., a mammalian cell, e.g., a human cell), the nucleic acid molecule may be cleaved into a dsRNA molecule wherein the two strands of the dsRNA molecule are no longer part of the same nucleic acid molecule e.g., by a Type III endonuclease (e.g., Dicer) (see, e.g., Sharp et al. (2001) Genes Dev. 15:485, the entire contents of which are incorporated herein by reference for all purposes).
(ii) Length of Double Stranded Region
[00262] In some embodiments, the double stranded region is about 15-30 base pairs in length (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17,
18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28,
19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25,
20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 base pairs in length). In some embodiments, the double stranded region is about 18-25 base pairs in length (e.g., 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20,
20-25, 20-24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 base pairs in length e.g., 19-21 base pairs in length)). In some embodiments, the double stranded region is about 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15- 20, 15-19, 15-18, 15-17, 15-16, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-20, 19-21, 23-30, 23-29, 23-28, 23-27, 23-26, 23-25, 23-24, 21-30, 21-29, 21-28, 21-27, 21-26,
21-25, 21-24, 21-23, or 21-22 base pairs in length. In some embodiments, the double stranded region is about 19-21 e.g., 19-20) base pairs in length.
[00263] In some embodiments, the double stranded region is not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length. In some embodiments, the double stranded region is about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length. In some embodiments, the double stranded region is about 19, 20, or 21 base pairs in length. In some embodiments, the double stranded region is about 19 base pairs in length. In some embodiments, the double stranded region is about 20 base pairs in length. In some embodiments, the double stranded region is about 21 base pairs in length. In some embodiments, the double stranded region is about 23 base pairs in length.
[00264] In some embodiments, the double stranded region comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length. In some embodiments, the double stranded region comprises about 19, 20, or 21 base pairs in length. In some embodiments, the double stranded region comprises about 19 base pairs in length. In some embodiments, the double stranded region comprises about 20 base pairs in length. In some embodiments, the double stranded region comprises about 21 base pairs in length. In some embodiments, the double stranded region is about 23 base pairs in length. [00265] In some embodiments, the double stranded region consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 base pairs in length. In some embodiments, the double stranded region consists of about 19, 20, or 21 base pairs in length. In some embodiments, the double stranded region consists of about 19 base pairs in length. In some embodiments, the double stranded region consists of about 20 base pairs in length. In some embodiments, the double stranded region consists of about 21 base pairs in length. In some embodiments, the double stranded region is about 23 base pairs in length.
[00266] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
(iii) Nucleotide Overhangs & Blunt Ends
[00267] In some embodiments, the dsRNA agent comprises one or more nucleotide overhang. As is clear- from the disclosure, but for the sake of clarity, the nucleotides of a nucleotide overhang can include one or more a modified (e.g., chemically modified) nucleotide (e.g., described herein, e.g., described in §§ 4.3, 4.3.1).
[00268] In some embodiments, the nucleotide overhang comprises from about 1-5 nucleotides, e.g., 1-4, 1-3, 1-2, 2-5, 2-4, 2-3, 3-5, 3-4, 4-5 nucleotides. In some embodiments, the nucleotide overhang comprises about 1, 2, 3, 4, or 5 nucleotides. In some embodiments, the nucleotide overhang comprises about 1 nucleotide. In some embodiments, the nucleotide overhang comprises about 2 nucleotides. In some embodiments, the nucleotide overhang consists of about 1, 2, 3, 4, or 5 nucleotides. In some embodiments, the nucleotide overhang consists of about 1 nucleotide. In some embodiments, the nucleotide overhang consists of about 2 nucleotides.
[00269] The nucleotide overhang(s) can be on the sense strand, the antisense strand, or both the sense strand and the antisense strand. In some embodiments, the sense strand comprises a nucleotide overhang. In some embodiments, the antisense strand comprises a nucleotide overhang. In some embodiments, the sense strand and the antisense strand both comprise a nucleotide overhang.
[00270] Furthermore, the nucleotide(s) of an overhang can be present on the 5'-end, 3'- end, or both the 5'-end, 3'- end of an antisense and/or sense strand. In some embodiments, the sense strand comprises a nucleotide overhang at the 5'-end. In some embodiments, the sense strand comprises a nucleotide overhang at the 3'-end. In some embodiments, the sense strand comprises a nucleotide overhang at the 5'-end and the 3 '-end. In some embodiments, the antisense strand comprises a nucleotide overhang at the 5'-end. Tn some embodiments, the antisense strand comprises a nucleotide overhang at the 3'-cnd. In some embodiments, the antisense strand comprises a nucleotide overhang at the 5'-end and the 3'-end. In some embodiments, the antisense strand comprises a nucleotide overhang at the 3'-end; and the sense strand comprises a nucleotide overhang at the 3'-end. In some embodiments, the antisense strand comprises a nucleotide overhang at the 5'-end; and the sense strand comprises a nucleotide overhang at the 5'-end.
[00271] In some embodiments, the dsRNA agent comprises one or more blunt end. In some embodiments, the dsRNA agent comprises a blunt end at the end of the RNA agent comprising the 3 'end of the sense strand and the 5' end of the antisense strand. In some embodiments, the dsRNA agent comprises a blunt end at the end of the RNA agent comprising the 5 'end of the sense strand and the 3' end of the antisense strand. In some embodiments, both ends of the dsRNA agent are blunt ends.
[00272] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
(iv) Exemplary Structural Combinations of Sense & Antisense Strands
[00273] In some embodiments, the antisense strand and the sense strand contain the same number of nucleotides. In some embodiments, the antisense strand and the sense strand contain different numbers of nucleotides. In some embodiments, the nucleotide sequence of the sense strand is from about 1-5, 1-3, or 1-2 nucleotides shorter than the nucleotide sequence of the antisense strand. In some embodiments, the nucleotide sequence of the sense strand is about 1, 2, 3, 4, or 5 nucleotides shorter than the nucleotide sequence of the antisense strand. In some embodiments, the nucleotide sequence of the sense strand is about 2 nucleotides shorter than the nucleotide sequence of the antisense strand. In some embodiments, the nucleotide sequence of the antisense strand is from about 1-5, 1-3, or 1-2 nucleotides shorter than the nucleotide sequence of the sense strand. In some embodiments, the nucleotide sequence of the antisense strand is about 1, 2, 3, 4, or 5 nucleotides shorter than the nucleotide sequence of the sense strand. In some embodiments, the nucleotide sequence of the antisense strand is about 2 nucleotides shorter than the nucleotide sequence of the sense strand.
[00274] In some embodiments, the sense strand comprises 21 nucleotides. In some embodiments, the antisense strand comprises 23 nucleotides. In some embodiments, the sense strand comprises 21 nucleotides; and the antisense strand comprises 23 nucleotides. In some embodiments, the double stranded region comprises 21 nucleotides. Tn some embodiments, the antisense strand comprises a 2-nuclcotidc overhang at the 3' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. In some embodiments, the sense strand comprises 21 nucleotides; the antisense strand comprises 23 nucleotides; the double stranded region comprises 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. [00275] In some embodiments, the sense strand comprises 19 nucleotides. In some embodiments, the antisense strand comprises 21 nucleotides. In some embodiments, the sense strand comprises 19 nucleotides; and the antisense strand comprises 21 nucleotides. In some embodiments, the double stranded region comprises 19 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. In some embodiments, the sense strand comprises 19 nucleotides; the antisense strand comprises 21 nucleotides; the double stranded region comprises 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. [00276] In some embodiments, the sense strand comprises 21 nucleotides. In some embodiments, the antisense strand comprises 21 nucleotides. In some embodiments, the sense strand comprises 21 nucleotides; and the antisense strand comprises 21 nucleotides. In some embodiments, the double stranded region comprises 19 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3'end. In some embodiments, the sense strand comprises a 2-nucleotide overhang at the 3'end. In some embodiments, the sense strand comprises 21 nucleotides; the antisense strand comprises 21 nucleotides; the double stranded region comprises 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 2-nucleotide overhang at the 3' end.
[00277] In some embodiments, the sense strand comprises 20 nucleotides. In some embodiments, the antisense strand comprises 19 nucleotides. In some embodiments, the sense strand comprises 20 nucleotides; and the antisense strand comprises 19 nucleotides. In some embodiments, the double stranded region comprises 20 nucleotides. In some embodiments, the sense strand comprises a 1 -nucleotide overhang at the 5' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. In some embodiments, the sense strand comprises 20 nucleotides; the antisense strand comprises 19 nucleotides; the double stranded region comprises 20 nucleotides; the sense strand comprises a 1 -nucleotide overhang at the 5' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
[00278] In some embodiments, the sense strand comprises 21 nucleotides. In some embodiments, the antisense strand comprises 19 nucleotides. In some embodiments, the sense strand comprises 21 nucleotides; and the antisense strand comprises 19 nucleotides. In some embodiments, the double stranded region comprises 19 nucleotides. In some embodiments, the sense strand comprises a 1 -nucleotide overhang at the 3' end. In some embodiments, the sense strand comprises a 1 -nucleotide overhang at the 5' end. In some embodiments, the sense strand comprises 21 nucleotides; the antisense strand comprises 19 nucleotides; the double stranded region comprises 19 nucleotides; the sense strand comprises a 1 -nucleotide overhang at the 3' end; and the sense strand comprises a 1 -nucleotide overhang at the 5' end.
[00279] In some embodiments, the sense strand comprises 24 nucleotides. In some embodiments, the antisense strand comprises 23 nucleotides. In some embodiments, the sense strand comprises 24 nucleotides; and the antisense strand comprises 23 nucleotides. In some embodiments, the double stranded region comprises 21 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3 'end. In some embodiments, the sense strand comprises a 3-nucleotide overhang at the 3' end. In some embodiments, the sense strand comprises 24 nucleotides; the antisense strand comprises 23 nucleotides; the double stranded region comprises 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 3-nucleotide overhang at the 3' end.
[00280] In some embodiments, the sense strand comprises 19 nucleotides. In some embodiments, the antisense strand comprises 19 nucleotides. In some embodiments, the sense strand comprises 19 nucleotides; and the antisense strand comprises 19 nucleotides. In some embodiments, the double stranded region comprises 19 nucleotides. In some embodiments, the 5' end of the antisense strand (and 3' end of the sense strand) form a blunt end. In some embodiments, the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end. In some embodiments, the sense strand comprises 19 nucleotides; the antisense strand comprises 19 nucleotides; the double stranded region comprises 19 nucleotides; the 5' end of the antisense strand (and 3' end of the sense strand) form a blunt end; and the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end
[00281] In some embodiments, the antisense strand and the sense strand are part of the same larger nucleic acid molecule, wherein the nucleic acid molecule comprises 44 nucleotides, the antisense portion comprises 21 nucleotides, the sense strand portion of the nucleic acid molecule comprises 19 nucleotides, the double stranded region comprises 19 nucleotides, the antisense strand comprises a 2-nucleotide overhang at the 3 'end, and the intervening nucleotide sequence between the antisense strand and the sense strand comprises 4 unpaired nucleotides that create a hairpin loop.
[00282] In some embodiments, the sense strand consists of 21 nucleotides. In some embodiments, the antisense strand consists of 23 nucleotides. In some embodiments, the sense strand consists of 21 nucleotides; and the antisense strand consists of 23 nucleotides. In some embodiments, the double stranded region consists of 21 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. In some embodiments, the sense strand consists of 21 nucleotides; the antisense strand consists of 23 nucleotides; the double stranded region consists of 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. [00283] In some embodiments, the sense strand consists of 19 nucleotides. In some embodiments, the antisense strand consists of 21 nucleotides. In some embodiments, the sense strand consists of 19 nucleotides; and the antisense strand consists of 21 nucleotides. In some embodiments, the double stranded region consists of 19 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3 'end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. In some embodiments, the sense strand consists of 19 nucleotides; the antisense strand consists of 21 nucleotides; the double stranded region consists of 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. [00284] In some embodiments, the sense strand consists of 21 nucleotides. In some embodiments, the antisense strand consists of 21 nucleotides. In some embodiments, the sense strand consists of 21 nucleotides; and the antisense strand consists of 21 nucleotides. In some embodiments, the double stranded region consists of 19 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3 'end. In some embodiments, the sense strand comprises a 2-nucleotide overhang at the 3'end. In some embodiments, the sense strand consists of 21 nucleotides; the antisense strand consists of 21 nucleotides; the double stranded region consists of 19 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 2-nuclcotidc overhang at the 3' end.
[00285] In some embodiments, the sense strand consists of 20 nucleotides. In some embodiments, the antisense strand consists of 19 nucleotides. In some embodiments, the sense strand consists of 20 nucleotides; and the antisense strand consists of 19 nucleotides. In some embodiments, the double stranded region consists of 20 nucleotides. In some embodiments, the sense strand comprises a 1-nucleotide overhang at the 5' end. In some embodiments, the 5' end of the antisense strand and 3' end of the sense strand form a blunt end. In some embodiments, the sense strand consists of 20 nucleotides; the antisense strand consists of 19 nucleotides; the double stranded region consists of 20 nucleotides; the sense strand comprises a 1-nucleotide overhang at the 5' end; and the 5' end of the antisense strand and 3' end of the sense strand form a blunt end.
[00286] In some embodiments, the sense strand consists of 21 nucleotides. In some embodiments, the antisense strand consists of 19 nucleotides. In some embodiments, the sense strand consists of 21 nucleotides; and the antisense strand consists of 19 nucleotides. In some embodiments, the double stranded region consists of 19 nucleotides. In some embodiments, the sense strand comprises a 1-nucleotide overhang at the 3' end. In some embodiments, the sense strand comprises a 1-nucleotide overhang at the 5' end. In some embodiments, the sense strand consists of 21 nucleotides; the antisense strand consists of 19 nucleotides; the double stranded region consists of 19 nucleotides; the sense strand comprises a 1-nucleotide overhang at the 3' end; and the sense strand comprises a 1-nucleotide overhang at the 5' end.
[00287] In some embodiments, the sense strand consists of 24 nucleotides. In some embodiments, the antisense strand consists of 23 nucleotides. In some embodiments, the sense strand consists of 24 nucleotides; and the antisense strand consists of 23 nucleotides. In some embodiments, the double stranded region consists of 21 nucleotides. In some embodiments, the antisense strand comprises a 2-nucleotide overhang at the 3'end. In some embodiments, the sense strand comprises a 3-nucleotide overhang at the 3'end. In some embodiments, the sense strand consists of 24 nucleotides; the antisense strand consists of 23 nucleotides; the double stranded region consists of 21 nucleotides; the antisense strand comprises a 2-nucleotide overhang at the 3' end; and the sense strand comprises a 3-nucleotide overhang at the 3' end.
[00288] In some embodiments, the sense strand consists of 19 nucleotides. In some embodiments, the antisense strand consists of 19 nucleotides. In some embodiments, the sense strand consists of 19 nucleotides; and the antisense strand consists of 19 nucleotides. In some embodiments, the double stranded region consists of 19 nucleotides. In some embodiments, the 5' end of the antisense strand (and 3’ end of the sense strand) form a blunt end. In some embodiments, the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end. In some embodiments, the sense strand consists of 19 nucleotides; the antisense strand consists of 19 nucleotides; the double stranded region consists of 19 nucleotides; the 5' end of the antisense strand (and 3' end of the sense strand) form a blunt end; and the 3' end of the antisense strand (and 5' end of the sense strand) form a blunt end
[00289] In some embodiments, the antisense strand and the sense strand are part of the same larger nucleic acid molecule, wherein the nucleic acid molecule consists of 44 nucleotides, the antisense portion consists of 21 nucleotides, the sense strand portion of the nucleic acid molecule comprises 19 nucleotides, the double stranded region consists of 19 nucleotides, the antisense strand comprises a 2-nucleotide overhang at the 3 'end, and the intervening nucleotide sequence between the antisense strand and the sense strand consists of 4 unpaired nucleotides that create a hairpin loop.
(v) Exemplary Antisense Strands & Sense Strands
[00290] In some embodiments, the antisense strand is an antisense strand described herein. In some embodiments, the sense strand is a sense strand described herein. In some embodiments, the antisense strand is an antisense strand described in § 4.2.1.1. In some embodiments, the sense strand is a sense strand described in § 4.2.1.2. In some embodiments, the antisense strand is an antisense strand described in § 4.2.1.1; and the sense strand is a sense strand described in § 4.2.1.2. It is to be understood that any sense strand described herein (e.g., in § 4.2.1.2); and be utilized in combination with any antisense strand in a dsRNA agent described herein (e.g., in § 4.2.1.1). For the sake of clarity, the entire contents of in §§ 4.2.1.1 and § 4.2.1.2, are incorporated by reference into the instant section.
(vi) Exemplary dsRNA Agents
[00291] The nucleotide sequence of exemplary unmodified and/or modified dsRNAi agents comprising a sense and antisense strand (e.g., suitable for targeting BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6, suitable for inhibiting BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 expression)) are described in herein.
[00292] In some embodiments, the dsRNA agent (e.g., for inhibiting expression of BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 (e.g., human BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of any one of SEQ ID NOS: 1, 4, 7, 10, 13, or 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17.
[00293] In some embodiments, the dsRNA agent (e.g., for inhibiting expression of BTG1 (e.g., (hBTGl) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding BTG1 (e.g., (hBTGl), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0,
1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 2. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1,
2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 1. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
[00294] In some embodiments, the dsRNA agent (e.g., for inhibiting expression of CDKN1B (e.g., (hCDKNIB) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding CDKN1B (e.g., (hCDKNIB), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 5. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1 , 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
[00295] In some embodiments, the dsRNA agent (e.g., for inhibiting expression of DUSP2 (e.g., (hDUSP2) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding DUSP2 (e.g., (hDUSP2), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0,
1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 8. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1,
2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
[00296] In some embodiments, the dsRNA agent (e.g., for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 11. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 10. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
[00297] In some embodiments, the dsRNA agent (e.g., for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 14. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 13. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
[00298] In some embodiments, the dsRNA agent (e.g., for inhibiting expression of KLHL6 (e.g., (hKLHL6) comprises a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding KLHL6 (e.g., (hKLHL6), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 17. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
[00299] Various salts, mixed salts and free acid forms of the dsRNA agents are also provided herein. In some embodiments, the dsRNA agent is in a free acid form. In some embodiments, the dsRNA agent is in a salt form. In some embodiments, the dsRNA agent is in a sodium salt form. In some embodiments, wherein the dsRNA agent is in the sodium salt form, sodium ions are present in the composition comprising the dsRNA agent as counterions for substantially all of the phosphodiester or phosphoro thioate groups present in the dsRNA agent. In some embodiments, wherein the dsRNA agent is in the sodium salt form, sodium ions are present in the RNA agent as counterions for all of the phosphodiester or phosphorothioate groups present in the dsRNA agent.
4.2.2 Antisense Oligonucleotide RNA Agents [00300] In some embodiments, the RNA agent comprises an antisense oligonucleotide. In some embodiments, the antisense oligonucleotide is single stranded. In some embodiments, the antisense oligonucleotide is single stranded and is not part of an RNAi agent.
[00301] Antisense oligonucleotides can mediate the inhibition or alteration of gene expression through, e.g., degradation of a target RNA (e.g., through the recruitment of endogenous enzymes (e.g., RNAses), inhibition of translation of a target RNA (e.g., through steric hinderance), and splicing modulation (e.g., leading to exon skipping, exon inclusion, etc.). See, e.g., Collotta, D et al. “Antisense oligonucleotides: a novel Frontier in pharmacological strategy.” Frontiers in pharmacology vol. 14 1304342. 17 Nov. 2023, doi:10.3389/fphar.2023.1304342, the entire contents of which are incorporated herein by reference for all purposes.
4.2.2.1 Targeting Region
[00302] As described herein, antisense oligonucleotides (e.g., described herein) comprise a region of complementarity that comprises a nucleotide sequence that is at least partially (e.g., substantially, fully) complementary to the nucleotide sequence of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is at least substantially complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is fully complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
[00303] In some embodiments, the nucleotide sequence of the region of complementarity is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, 1GLL5, or KLHL6 mRNA)). For example, the nucleotide sequence of the region of complementarity may be at least 70% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., aBTGl, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 75% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 80% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 85% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 90% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). The nucleotide sequence of the region of complementarity may be at least 95% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is at least 95%, 96%, 97%, 98%, 99%, or 100% (e.g., in some embodiments, preferably at least 95%, more preferably at least 98%) complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity is 100% complementary to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)).
[00304] In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of one or more non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)). In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 5 e.g., 4, 3, 2, 1, or 0) non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 3 (e.g., 2, 1, or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises or consists of no more than 2 (e.g., 1 or 0) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises no more than 1 (e.g., 0) non-complementary nucleotide mismatch relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the nucleotide sequence of the region of complementarity comprises 0 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule. In some embodiments, the region of complementarity comprises one or more (e.g., 2, 3, or more) non- complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity. In some embodiments, the region of complementarity comprises at least one but not more than 3 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 5 (e.g., 4, 3, 2, or 1) nucleotides from either the 5'- and/or 3'-end of the region of complementarity. In some embodiments, the region of complementarity comprises one or more (e.g., 2, 3, or more) non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non- complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity. In some embodiments, the region of complementarity comprises at least one but not more than 3 non-complementary nucleotide mismatches relative to the nucleotide sequence of the target nucleic acid molecule, wherein the one or more non-complementary nucleotide mismatches are within the last 3 (e.g., 2 or 1) nucleotides from either the 5'- and/or 3 '-end of the region of complementarity. Methods known in the art and described herein can be utilized to evaluate the effect of any non-complementary mismatches between an antisense strand and a target nucleic acid molecule on functional properties (e.g., inhibition of expression of the target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), a portion of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA))).
[00305] In some embodiments, the region of complementarity comprises from about 15-30 nucleotides, e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30,
19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27,
20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides. In some embodiments, the region of complementarity comprises from about 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-
24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides. In some embodiments, the region of complementarity comprises from about 19-21 (e.g., 19-20) nucleotides. In some embodiments, the region of complementarity comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity comprises about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity comprises about 19 nucleotides. In some embodiments, the region of complementarity comprises about 20 nucleotides. In some embodiments, the region of complementarity comprises about 21 nucleotides. In some embodiments, the region of complementarity comprises about 22 nucleotides. In some embodiments, the region of complementarity comprises about 23 nucleotides. In some embodiments, the region of complementarity consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the region of complementarity consists of about 19, 20, 21, 22, or 23 nucleotides. In some embodiments, the region of complementarity consists of about 19 nucleotides. In some embodiments, the region of complementarity consists of about 20 nucleotides. In some embodiments, the region of complementarity consists of about 21 nucleotides. In some embodiments, the region of complementarity consists of about 22 nucleotides. In some embodiments, the region of complementarity consists of about 23 nucleotides. Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
[00306] In some embodiments, the target nucleic acid molecule is part (e.g., a contiguous portion) of a larger nucleic acid molecule. For example, in some embodiments, the target nucleic acid molecule is a portion (e.g., a contiguous portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA). In some embodiments, the target nucleic acid molecule is a contiguous nucleotide sequence of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) of sufficient length to allow it to be a substrate for cleavage directed by the antisense oligonucleotide.
[00307] In some embodiments, the target nucleic acid molecule is a target mRNA (e.g., a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA). In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA). In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA) formed in the expression of a target gene (e.g., a mammalian, primate, human, non-human primate, mouse, and/or rat gene) (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 gene).
[00308] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a BTG1 (e.g., hBTGl) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a BTG1 (e.g., hBTGl) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 1 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a BTG1 (e.g., hBTGl) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 1 (or a variant or fragment thereof).
[00309] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a CDKN1B (e.g., hCDKNIB) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a CDKN 1 B (e.g., hCDKNIB) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 4 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a CDKN1B (e.g., hCDKNIB) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 4 (or a variant or fragment thereof).
[00310] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a DUSP2 (e.g., hDUSP2) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a DUSP2 (e.g., hDUSP2) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 7 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a DUSP2 (e.g., hDUSP2) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 7 (or a variant or fragment thereof).
[00311] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a IGLL5 (e.g., hIGLL5) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a IGLL5 (e.g., hIGLL5) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 10 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 10 (or a variant or fragment thereof). [00312] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a IGLL5 (e.g., hIGLL5) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a IGLL5 (e.g., hIGLL5) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 13 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a IGLL5 (e.g., hIGLL5) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 13 (or a variant or fragment thereof).
[00313] In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of a KLHL6 (e.g., hKLHL6) mRNA. In some embodiments, the target nucleic acid molecule is at least a portion (e.g., a portion) of an mRNA formed in the expression of a KLHL6 (e.g., hKLHL6) gene. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of the nucleotide sequence set forth in SEQ ID NO: 16 (or a variant or fragment thereof). In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a target protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA encoding a KLHL6 (e.g., hKLHL6) protein. In some embodiments, the target nucleic acid molecule comprises at least a portion (e.g., a portion) of an mRNA sequence encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 16 (or a variant or fragment thereof).
[00314] In some embodiments, the target nucleic acid molecule comprises from about 19-30 nucleotides, e.g., 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20- 29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, 21-22, 22-30, 22-29, 22-28, 22-27, 22-26, 22-25, 22-24, 22-23, 23-30, 23-29, 23-28, 23-27, 23-26, 23-27, 23-26, 23-25, or 23-24 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 19-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 19-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-25 nucleotides. In some embodiments, the target nucleic acid molecule comprises from about 21-23 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 19, 18, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the target nucleic acid molecule comprises or consists of about 19 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 20 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 21 nucleotides. In some embodiments, the target nucleic acid molecule comprises about 23 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 19, 18, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 19 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 20 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 21 nucleotides. In some embodiments, the target nucleic acid molecule consists of about 23 nucleotides.
[00315] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
4.2.2.2 Overall Length
[00316] In some embodiments, the antisense oligonucleotide comprises from about 15-50 nucleotides (e.g., 15-49, 15-48, 15-47, 15-46, 15-45, 15-44, 15-43, 15-42, 15-41, 15-40, 15-39, 15-38, 15-37, 15-36, 15-35, 15-34, 15-33, 15-32, 15-31, 15-30, 15-29, 15-28, 15-27, 15- 26, 15- 25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26,
18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23,
19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides).
[00317] In some embodiments, the antisense oligonucleotide comprises from about 15-35 nucleotides (e.g., 15-34, 15-33, 15-32, 15-31, 15-30, 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24,
15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24,
18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21,
19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21, 21-30, 21-29, 21-28,
21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides).
[00318] In some embodiments, the antisense oligonucleotide comprises from about 15-30 nucleotides (e.g., 15-29, 15-28, 15-27, 15- 26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24,20-23, 20-22, 20-21 , 21-30, 21 -29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides).
[00319] In some embodiments, the antisense oligonucleotide comprises from about 18-35 nucleotides (e.g., 18-34, 18-33, 18-32, 18-31, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides). In some embodiments, the antisense oligonucleotide comprises from about 18-30 nucleotides (e.g.,
18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22,
19-21, 19-20, 20-25, 20-24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25, 22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides). In some embodiments, the antisense oligonucleotide comprises from about 18-25 nucleotides (e.g., 18-24, 18-23, 18-22, 18-21, 18-20, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-25, 20-24,20-23, 20-22, 20-21, 21-25, 21-24, 21-23, 21-22, 22- 25,
22-24, 22-23, 23-25, 23-24 or 24-25 nucleotides). In some embodiments, the antisense oligonucleotide comprises from about 19-25 nucleotide (e.g., 19-20, 19-21, 19-22, 19-23, 19-24, 19-25, 20-21, 20-22, 20-23, 20-24, 20-25, 21-22, 21-23, 21-24, 21-25, 22-23, 22-24, 22-25, 23-24,
23-25, 24-25 nucleotides). In some embodiments, the antisense oligonucleotide comprises from about 15-30, 16-30, 17-30, 18-30, 19-3020-30, 21-30, 22-30, 23-30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides.
[00320] In some embodiments, the antisense oligonucleotide comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides. In some embodiments, the antisense oligonucleotide comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides. In some embodiments, the antisense oligonucleotide comprises not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
[00321] In some embodiments, the antisense oligonucleotide consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides. In some embodiments, the antisense oligonucleotide consists of not more than about 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides. In some embodiments, the antisense oligonucleotide consists of not more than about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.
[00322] In some embodiments, the antisense oligonucleotide comprises about 18 nucleotides.
In some embodiments, the antisense oligonucleotide comprises about 19 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 20 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 21 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 22 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 23 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 24 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 25 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 26 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 27 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 28 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 29 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 30 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 31 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 32 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 33 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 34 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 35 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 36 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 37 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 38 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 39 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 40 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 41 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 42 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 43 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 44 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 45 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 46 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 47 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 48 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 49 nucleotides. In some embodiments, the antisense oligonucleotide comprises about 50 nucleotides.
[00323] In some embodiments, the antisense oligonucleotide consists of about 18 nucleotides.
In some embodiments, the antisense oligonucleotide consists of about 19 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 20 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 21 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 23 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 22 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 23 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 24 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 25 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 26 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 27 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 28 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 29 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 30 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 31 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 32 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 33 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 34 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 35 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 36 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 37 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 38 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 39 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 40 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 41 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 42 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 43 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 44 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 45 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 46 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 47 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 48 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 49 nucleotides. In some embodiments, the antisense oligonucleotide consists of about 50 nucleotides.
[00324] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
4.2.2.3 Exemplary Antisense Oligonucleotides
[00325] In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides. [00326] In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00327] In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00328] In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31 , 32, 33, 34, or 35) nucleotides from the nucleotide sequence of a portion of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides. [00329] In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00330] In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00331] In some embodiments, the antisense oligonucleotide comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 19 contiguous nucleotides.
[00332] In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides.
[00333] In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides.
[00334] In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides.
[00335] In some embodiments, the antisense oligonucleotide comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide comprises 23 contiguous nucleotides.
[00336] In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00337] In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00338] In some embodiments, the antisense oligonucleotide consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 19 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 19 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 19 contiguous nucleotides.
[00339] In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides.
[00340] In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides.
[00341] In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides.
[00342] In some embodiments, the antisense oligonucleotide consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 23 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 23 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of the reverse complement of a portion of a BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6 mRNA. In some embodiments, the antisense oligonucleotide consists of 23 contiguous nucleotides.
[00343] In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 15 (e.g., at least 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00344] In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0,
I, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 20 (e.g., at least 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides.
[00345] In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8,
I I, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 25 (e.g., at least 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) contiguous nucleotides. [00346] In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises at least 30 (e.g., at least 31, 32, 33, 34, or 35) contiguous nucleotides.
[00347] In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,
25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00348] In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25,
26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00349] In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 20 contiguous nucleotides. [00350] In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 21 contiguous nucleotides.
[00351] In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 22 contiguous nucleotides.
[00352] In some embodiments, the antisense oligonucleotide comprises 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 30 contiguous nucleotides differing by no more than I nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide comprises 30 contiguous nucleotides.
[00353] In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,
25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00354] In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25,
26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35 contiguous nucleotides.
[00355] In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 20 contiguous nucleotides.
[00356] In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 21 contiguous nucleotides.
[00357] In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 22 contiguous nucleotides.
[00358] In some embodiments, the antisense oligonucleotide consists of 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1 , 2, 3, 4, or 5) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 30 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) (e.g., by no more than 3 (e.g., 0, 1, 2, or 3)) nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 30 contiguous nucleotides differing by no more than 1 nucleotides from the nucleotide sequence of any one of SEQ ID NOS: 2, 5, 8, 11, 14, or 17. In some embodiments, the antisense oligonucleotide consists of 30 contiguous nucleotides.
[00359] Ranges and lengths intermediate to the above recited ranges and lengths are also contemplated to be part of the disclosure.
4.3 Modified RNA Agents
[00360] In some embodiments, the RNA agent (e.g., RNAi agent (e.g., siRNA)) (or any component thereof (e.g., any nucleic acid molecule thereof)) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, antisense oligonucleotide, etc.) comprises one or more modified nucleotide(s) (as defined herein). The modified agent may have one or more different (e.g., improved) properties relative to a corresponding unmodified RNA agent. For example, the modified RNA agent may exhibit decreased immunostimulatory activity (e.g., when administered to a subject), increased stability (e.g., in vivo, in a cell, when administered to a subject), and/or increased inhibition of expression of a target nucleic acid molecule (e.g., a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA)), or any combination thereof.
4.3.1 Nature of Nucleotide Modifications
[00361] Nucleotide modifications can include modification to any one of more of the nucleoside and/or the internucleoside linkage. Nucleoside modifications include modification to the sugar (e.g., ribose) moiety and/or the nucleobase. In some embodiments, the modified agent (or component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more nucleotides comprising a modified sugar moiety. In some embodiments, the modified agent (or component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more nucleotides comprising a modified nucleobase. In some embodiments, the modified agent (or component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more nucleotides comprising a modified intemucleoside linkage. In some embodiments, the modified agent (or component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more nucleotides comprising one, two, or three of a modified sugar moiety, a modified nucleobase, and/or a modified intemucleoside linkage.
[00362] Exemplary nucleotide modifications are described below and also known in the art, see, e.g., WO2021257782, W02013075035, WO2022246251, and WO2022271573, the entire contents of each of which are incorporated by reference herein for all purposes.
4.3.1.1 Modified Nucleosides
[00363] In some embodiments, the modified RNA agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, antisense oligonucleotide, etc.) comprises one or more nucleotide comprising a modified nucleoside. As discussed above, nucleoside modifications can include modification of the sugar (e.g., ribose) moiety and/or modification of the nucleobase.
(i) Sugar Modifications
[00364] In some embodiments, the modified RNA agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, antisense oligonucleotide, etc.) comprises one or more nucleotides comprising a modified sugar (e.g., ribose) moiety.
[00365] The modified sugar (e.g., ribose) moiety can comprise, for example, a substituent at any one or more position of the sugar (e.g., ribose), including e.g., positions 2', 4', and/or 5'. In some embodiments, the modified sugar (e.g., ribose) comprises a substituent at 2' position of the sugar (e.g., ribose). In some embodiments, the modified sugar (e.g., ribose) comprises a substituent at 5' position of the sugar (e.g., ribose). In some embodiments, the modified sugar (e.g., ribose) comprises a substituent at 5' position of the sugar (e.g., ribose).
[00366] In some embodiments, the RNA agent (or any component thereof) comprises any one or more of the following substituents (e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')): a group for improving the stability of the RNA agent, a group for improving the pharmacokinetic properties of the RNA agent, or a group for improving the pharmacodynamic properties of the RNA agent, an RNA cleaving group, a reporter group, an intercalator, and/or other substituents having similar properties.
[00367] Exemplary substituents include, for example, but are not limited to, substitution (e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')) with any one of the following: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl can be substituted or unsubstituted Ci to Cio alkyl or C2 to Cio alkenyl and alkynyl. Additional exemplary substitutions (e.g., at any position of the sugar (e.g., ribose) (e.g., at position 2')) include, for example, but are not limited to, substitution with any one of the following: 0[(CH2)n0]m, CH3, O(CH2)UOCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, and O(CH2)nON[(CH2)nCH3)]2, where n and m are from 1 to about 10.
[00368] In some embodiments, the modified sugar (e.g., ribose) comprises any one of the following modifications: 2'-O-methyl (2'-0Me), 2'O-methoxyethyl (2'-0-M0E), 2'deoxy-2'- fluoro (2'-F), 2'-arabino-fluoro (2'-Ara-F), 2'-O-benzyl, 2'-O-methyl-4-pyridine (2-O-methyl-4- pyridine (2'-O-CH2Py(4)).
[00369] In some embodiments, the RNA agent (or any component thereof) comprises any of the following substituents at the 2'-position of the sugar (e.g., ribose): Ci to Cio lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, or a substituted silyl. In some embodiments, the RNA agent (or any component thereof) comprises a 2'-methoxyethoxy (2'-0 — CH2CH2OCH3, also known as 2'-O-(2-mcthoxycthyl) or 2'-M0E) (see, e.g., Martin ct al., Hclv. Chim. Acta, 1995, 78:486-504, the entire contents of which is incorporated by reference herein for all purposes) (i.e., an alkoxyalkoxy group). In some embodiments, the RNA agent (or any component thereof) comprises a 2'- dimethylaminooxy ethoxy, i.e., a O(CH2)2ON(CH3)2 group, also known as 2'-DMAOE; a 2'- dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethylaminoethoxyethyl or 2'- DMAEOE), i.e., 2'-O— CH2— O— CH2— N(CH3)2; a 5'-Me-2'-F nucleotide, a 5'-Me-2'-OMe nucleotide, a 5'-Me-2'-deoxynucleotide, (both R and S isomers in these three families); a 2'- alkoxyalkyl; and 2'-NMA (N-methylacetamide).
[00370] Exemplary US patents that describe the preparation of such modified sugar structures include, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920; the entire contents of each of the foregoing are hereby incorporated herein by reference for all purposes.
(a) Non-Bicyclic Sugar' Modifications
[00371] In some embodiments, the modified sugar (e.g., ribose) moiety comprises a non- bicyclic modified sugar (e.g., ribose) moiety. In some embodiments, the modified sugar (e.g., ribose) moiety comprises a furanosyl ring comprising one or more substituent groups none of which bridges two atoms of the furanosyl ring to form a bicyclic structure. In some embodiments one or more non-bridging substituent of a non-bicyclic modified sugar moiety is branched. Such non bridging substituents may be at any position of the furanosyl, including but not limited to substituents at the 2', 4', and/or 5' positions.
[00372] In some embodiments, non-bicyclic modified sugar moiety comprises a substituent group at the 2'-position of the sugar (e.g., ribose). Examples of 2'- substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 2'-O-methyl (2'-0Me), 2'0- methoxyethyl (2'-0-M0E), 2 'deoxy-2 '-fluoro (2'-F), 2'-arabino-fluoro (2'-Ara-F), 2'-O-benzyl, 2'- O-methyl-4-pyridine (2-O-methyl-4-pyridine (2'-O-CH2Py(4)), and 2’-O-N-alkyl acetamide (e.g., 2'-O-N-methyl acetamide (“NMA”), 2'-O-N-dimethyl acetamide, 2'-O-N-ethyl acetamide, and 2'- O-N-propyl acetamide). For example, see, e.g., U.S. 6,147,200, Prakash et al., 2003, Org. Lett., 5, 403-6, the entire contents of which is incorporated by reference herein for all purposes. [00373] In some embodiments, the 2'-substituent group is a halo, allyl, amino, azido, SH, CN, OCN, CF3, OCF3, O-C1-C10 alkoxy, O-C1-C10 substituted alkoxy, O-C1-C10 alkyl, O-C1-C10 substituted alkyl, S-alkyl, N(Rm)-alkyl, O-alkenyl, S-alkenyl, N(Rm)-alkenyl, O-alkynyl, S- alkynyl, N(Rm)-alkynyl, O-alkylenyl-O- alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, O(CH2)2SCH3,0(CH2)2ON(Rm)(Rn) or OCH2C(=O)- N(Rm)(Rn), where each Rm and Rn is, independently, H, an amino protecting group, or substituted or unsubstituted C1-C10 alkyl, or a 2'- substituent group described in any one of the following: Cook et al., U.S. 6,531,584; Cook et al., U.S. 5,859,221; and Cook et al., U.S. 6,005,087, the entire contents of which are incorporated herein by reference for all purposes. In some embodiments, these 2'-substituent groups can be further substituted with one or more substituent groups independently selected from among: hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO2), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl.
[00374] In some embodiments, a 2'-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2'-substituent group selected from: F, NH2, N3, OCF3, 0CH3, O(CH2)3NH2, CH2CH=CH2, OCH2CH=CH2, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(Rm)(Rn), O(CH2)2O(CH2)2N(CH3)2, and N-substituted acetamide (OCH2C(=O)- N(Rm)(Rn)), where each Rm and Rn is, independently, H, an amino protecting group, or substituted or unsubstituted C1-C10 alkyl. In some embodiments, a 2 '-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2'-substituent group selected from: F, OCF, OCH3, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2) ON(CH3)2, O(CH2)2O(CH2)2N(CH3)2, and OCH2C(=O)-N(H)CH3 (“NMA”). In some embodiments, a 2'- substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2'-substituent group selected from: F, OCH3, OCH2CH2OCH3, and OCH2C(=O)-N(H)CH .
[00375] In some embodiments, non-bicyclic modified sugar moiety comprises a substituent group at the 3'-position of the sugar (e.g., ribose). Examples of substituent groups suitable for the 3 '-position of modified sugar moieties include but are not limited to alkoxy (e.g., methoxy), alkyl (e.g., methyl, ethyl).
[00376] In some embodiments, non-bicyclic modified sugar moiety comprises a substituent group at the 4'-position of the sugar (e.g., ribose). Examples of 4'-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to alkoxy (e.g., methoxy), alkyl, and those described in Manoharan et al., WO 2015/106128. [00377] In some embodiments, non-bicyclic modified sugar moiety comprises a substituent group at the 5 '-position of the sugar (e.g., ribose). Examples of substituent groups suitable for the 5'-position of modified sugar moieties include, but are not limited to, vinyl (e.g., 5'-vinyl), alkoxy (e.g., methoxy (e.g., 5'-methoxy)), and alkyl (e.g., methyl (R or S) (e.g., 5'-methyl (R or S)), ethyl). [00378] In some embodiments, non-bicyclic modified sugar moieties comprise more than one non-bridging sugar substituent, for example, 2'-F-5'-methyl sugar moieties and the modified sugar moieties and modified nucleosides described in Migawa et al., WO 2008/101157 and Rajeev et al., US2013/0203836, the entire contents of each of which is incorporated herein by reference for all purposes.
[00379] In some embodiments, modified furanosyl sugar moieties and nucleosides incorporating such modified furanosyl sugar moieties are further defined by isomeric configuration. For example, a 2'- deoxyfuranosyl sugar moiety may be in seven isomeric configurations other than the naturally occurring -D- deoxyribosyl configuration. Such modified sugar moieties are described in, e.g., WO 2019/157531, the entire contents of which are incorporated by reference herein for all purposes.
[00380] In some embodiments, the sugar (e.g., ribose) modification comprises an unlocked nucleotide (UNA). UNA is unlocked acyclic nucleic acid, wherein any of the bonds of the sugar has been removed, forming an unlocked sugar (e.g., ribose) residue. For example, in some embodiments, the bonds between Cl'-C4' have been removed (i.e. the covalent carbon-oxygen- carbon bond between the Cl' and C4' carbons). In some embodiments, the C2'-C3' bond (i.e. the covalent carbon-carbon bond between the C2' and C3' carbons) of the sugar (e.g., ribose) have been removed. See, e.g., Nuc. Acids Symp. Series, 52, 133-134 (2008) and Fluiter et al., Mol. Biosyst., 2009, 10, 1039, the entire contents of which are incorporated herein by reference. UNAs and methods of making are known in the art. See, e.g., U.S. Pat. No. 8,314,227; and US2013/0096289; US2013/0011922; and US2011/0313020, the entire contents of each of which are hereby incorporated herein by reference.
(b) Bicyclic Sugar Modifications
[00381] In some embodiments, the modified sugar’ (e.g., ribose) moiety comprises a substituent that bridges two atoms of the furanosyl ring to form a second ring, resulting in a bicyclic sugar (e.g., ribose) moiety. In some embodiments, the bicyclic sugar (e.g., ribose) moiety comprises a bridge between the 4’ and the 2’ furanose ring atoms. Examples of such 4' to 2' bridging sugar substituents include but are not limited to: 4'-CH2-2', 4'-(CH2)2-2', 4'-(CH2)3-2', 4'-CH2-O-2' (“LNA”), 4'-CH2-S-2’, 4'-(CH2)2-O-2' (“ENA”), 4'-CH(CH3)-O-2' (referred to as “constrained ethyl” or “cEt”), 4'-CH2- O-CH2-2', 4'-CH2-N(R)-2', 4'-CH(CH2OCH3)-O-2' (“constrained MOE” or “cMOE”) and analogs thereof (see, e.g., Seth et al., U.S. 7,399,845, Bhat et al., U.S. 7,569,686, Swayze et al., U.S. 7,741,457, and Swayze et al., U.S. 8,022,193), 4'-C(CH3)(CH3)-O-2' and analogs thereof (see, e.g., Seth et al., U.S. 8,278,283), 4'- CH2-N(OCH3)-2' and analogs thereof (see, e.g., Prakash et al., U.S. 8,278,425), 4'-CH2-O-N(CH3)-2' (see, e.g., Allerson et al., U.S. 7,696,345 and Allerson et al., U.S. 8,124,745), 4'-CH -C(H)(CH3)-2' (see, e.g., Zhou, et al., J. Org. Chem.,2QQ9, 74, 118-134), 4'-CH2-C(=CH2)-2' and analogs thereof (see, e.g., Seth et al., U.S. 8,278,426), 4'-C(RaRb)-N(R)-O-2', 4'-C(RaRb)-O-N(R)-2', 4'-CH2-O-N(R)-2’, and 4'-CH2-N(R)- 0-2', wherein each R, Ra, and Rb is, independently, H, a protecting group, or C1-C12 alkyl (see, e.g. Imanishi et al., U.S. 7,427,672). The entire contents of all of the foregoing references is incorporated by reference herein for all purposes.
[00382] In some embodiments, such 4' to 2' bridges independently comprise from 1 to 4 linked groups independently selected from: -[C(Ra)(Rb)]n-, -[C(Ra)(Rb)]n-0-, -C(Ra)=C(Rb)-, -C(Ra)=N-, -C(=NRa)-, -C(=O)-, -C(=S)-, -O-, -Si(Ra)2-, -S(=O)X-, and -N(Ra)-; wherein: x is 0, 1, or 2; n is 1, 2, 3, or 4; each Ra and Rb is, independently, H, a protecting group, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-Ci2 alkenyl, C2-Ci2 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)- H), substituted acyl, CN, sulfonyl (S(=0)2-Jl), or sulfoxyl (S(=O)-J1); and each JI and J2 is, independently, H, C1-C12 alkyl, substituted C1-C12 alkyl, C2-Ci2 alkenyl, substituted C2-Ci2 alkenyl, C2-Ci2 alkynyl, substituted C2-Ci2 alkynyl, C5-C20 aryl, substituted Cs-C2o aryl, acyl (C(=O)- H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C1-C12 aminoalkyl, substituted C1-C12 aminoalkyl, or a protecting group.
[00383] Additional bicyclic sugar moieties are known in the art, see, for example: Freier el al., Nucleic Acids Research, 1997, 25(22), 4429-4443, Albaek et al., J. Org. Chem., 2006, 71, 7731- 7740, Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J. Org. Chem., 1998, 63, 10035-10039; Srivastavaet al., J. Am. Chem. Soc., 2007, 129, 8362-8379;Wengel et a., U.S. 7,053,207; Imanishi et al., U.S. 6,268,490; Imanishi et al. U.S. 6,770,748; Imanishi et al., U.S. RE44,779; Wcngcl ct al., U.S. 6,794,499; Wcngcl ct al., U.S. 6,670,461; Wcngcl ct al., U.S. 7,034,133; Wengel et al., U.S. 8,080,644; Wengel et al., U.S. 8,034,909; Wengel et al., U.S. 8,153,365; Wengel et al., U.S. 7,572,582; Ramasamy et al., U.S. 6,525,191; Torsten et al., WO 2004/106356; Wengel et al., WO 1999/014226; Seth et al., WO 2007/134181; Seth et al., U.S. 7,547,684; Seth et al., U.S. 7,666,854; Seth et al., U.S. 8,088,746; Seth et al., U.S. 7,750,131; Seth et al., U.S. 8,030,467; Seth et al., U.S. 8,268,980; Seth et al., U.S. 8,546,556; Seth et al., U.S. 8,530,640; Migawa et al., U.S. 9,012,421; Seth et al., U.S. 8,501,805; and U.S. Patent Publication Nos. Allerson et al., US2008/0039618 and Migawa et al., US2015/0191727. The entire contents of all of the foregoing references is incorporated by reference herein for all purposes.
[00384] In some embodiments, the modified sugar (e.g., ribose) comprises a constrained ethyl nucleotide comprising a 4'-CH(CH3) — O-2' bridge. In some embodiments, the constrained ethyl nucleotide is in the S conformation (S-cEt). In some embodiments, the modified sugar (e.g., ribose) comprises a conformationally restricted nucleotide (CRN). CRNs are nucleotide analogs with a linker connecting the C2' and C4' carbons of ribose or the C3 and — C5' carbons of ribose. Representative publications that teach the preparation of certain of the above include, but are not limited to, US2013/0190383; and WO2013/036868, the entire contents of each of which are hereby incorporated herein by reference.
[00385] In some embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by isomeric configuration. For example, an LNA nucleoside (described herein) may be in the a-L configuration or in the 0-D configuration. Herein, general descriptions of bicyclic nucleosides include both isomeric configurations. Any of the foregoing bicyclic nucleosides can be prepared having one or more stereochemical sugar configurations including for example a-L-ribofuranose and -D-ribofuranose (see, e.g., WO 99/14226, the entire contents of which are incorporated herein by reference for all purposes).
[00386] Additional representative U.S. Patents and U.S. Patent Publications that teach the preparation of bicyclic nucleosides (e.g., locked nucleic acid) include, but arc not limited to, the following: U.S. Pat. Nos. 6,268,490; 6,525,191; 6,670,461; 6,770,748; 6,794,499; 6,998,484; 7,053,207; 7,034,133; 7,084,125; 7,399,845; 7,427,672; 7,569,686; 7,741,457; 8,022,193; 8,030,467; 8,278,425; 8,278,426; 8,278,283; US 2008/0039618; and US 2009/0012281, the entire contents of each of which are hereby incorporated herein by reference. (ii) Nucleobase Modifications
[00387] In some embodiments, the modified agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more nucleotides comprising a modified nucleobase.
[00388] As used herein, “unmodified” nucleobases refer to the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Modified nucleobases include other synthetic and natural nucleobases.
[00389] Modified nucleobases include, but are not limited to, 5-substituted pyrimidines, 6- azapyrimidines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, and N-2, N-6 and 0-6 substituted purines. In certain embodiments, modified nucleobases are selected from: 5- methylcytosine, 2-aminopropyladenine, 5 -hydroxymethyl cytosine, xanthine, hypoxanthine, deoxythimidine (dT), 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2- propyladenine , 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (-C=C-CH3) uracil, 5- propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4- thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, particularly 5-bromo, 5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7- methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N- benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-Nbenzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines, such as 1,3-diazaphenoxazine- 2-one, l,3-diazaphenothiazine-2-one and 9-(2-aminoethoxy)-l,3-diazaphenoxazine-2-one (G- clamp). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza- adenine, 7-deazaguanosine, 2- aminopyridine and 2-pyridone. Further nucleobases include those disclosed in Merigan et al., U.S. 3,687,808; The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; Sanghvi, Y.S., Chapter 15, Antisense Research and Applications, Crooke, S.T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6 and 15, Antisense Drug Technology, Crooke S.T., Ed., CRC Press, 2008, 163-166 and 442-443; the entire contents of each of which are incorporated herein by reference for all purposes. [00390] In some embodiments, the modified nucleobase comprises a pseudouridine, 2'thiouridinc (s2U), N6'-mcthyladcnosinc, 5 'mcthylcy tidinc (m5C), 5 'fluoro-2 'deoxy uridine, N- ethylpiperidine 7-EAA triazole modified adenine, N-ethylpiperidine 6'triazole modified adenine, 6-phenylpyrrolo-cytosine (PhpC), 2',4'-difluorotoluyl ribonucleoside (rF), or 5 'nitroindole. In some embodiments, the modified nucleobase comprises a 5-substituted pyrimidine; 6- azapyrimidine; or N-2, N-6 and 0-6 substituted purines (including 2-aminopropyladenine, 5- propynyluracil and 5-propynylcytosine). 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., Eds., dsRNA Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are exemplary base substitutions, even more particularly when combined with 2'-O-methoxyethyl sugar modifications.
[00391] Representative U.S. Patents an published applications that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include, but are not limited to, U.S. Pat. Nos. 3,687,808, 4,845,205; 5,130,30; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540;
5,587,469; 5,594,121, 5,596,091; 5,614,617; 5,681,941; 5,750,692; 6,015,886; 6,147,200;
6,166,197; 6,222,025; 6,235,887; 6,380,368; 6,528,640; 6,639,062; 6,617,438; 7,045,610;
7,427,672; 7,495,088; 5,130,302; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,434,257;
5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,711; 5,552,540; U.S. 5,587,469; 5,594,121; 5,596,091; 5,614,617; 5,645,985; 5,681,941; 5,811,534; 5,750,692; 5,948,903; 5,587,470;
5,457,191; 5,763,588; 5,830,653; 5,808,027; 6,166,199; and 6,005,096, the entire contents of each of which are hereby incorporated herein by reference for all purposes.
4.3.1.2 Internucleoside Linkage Modifications
[00392] In some embodiments, the modified agent (or any component thereof) (e.g., antisense strand, sense strand, dsRNA agent, etc.) comprises one or more modified internucleoside linkage. Modified internucleoside linkages, compared to naturally occurring phosphate linkages, can be used to alter, typically increase, nuclease resistance of an RNA agent (e.g., described herein).
[00393] The naturally occurring internucleoside linkage of RNA and DNA is a 3’ to 5' phosphodiester linkage. In some embodiments, the modified internucleoside linkage contains a normal 3'-5' linkage. In some embodiments, the modified intemucleoside linkage contains a 2'-5' linkage. In some embodiments, the modified intemucleoside linkage has an inverted polarity wherein the adjacent pairs of nucleoside units are linked e.g., 3'-5' to 5'-3' or 2'- 5 ' to 5'-2'.
[00394] The two main classes of modified intcmuclcosidc linking can be defined by the presence or absence of a phosphorous atom.
(i) Modified Phosphorous Containing Internucleoside Linkages
[00395] In some embodiments, the modified intemucleoside linkage comprises a phosphorous atom. Representative modified phosphorus-containing internucleoside linkages include but are not limited to phosphorothioates (PS (Rp isomer or Sp isomer)) (e.g., 5'phosphorothioate), phosphotriesters, phosphoramidates e.g., 3'-amino phosphoramidate and aminoalky Iphosphoramidates), chiral phosphorothioates, phosphorodithioates (PS2), aminoalkylphosphotriesters, methyl and other alkyl phosphonates (e.g., methylphosphonate (MP), 3'-alkylene phosphonates), methpxypropyl-phosphonates (MOP), 5'-(E)-vinylphosphonates, 5'methyl phosphonates, (S)-5'C-methyl with phosphates, phosphinates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, boranophosphates, and peptide nucleic acids (PNAs).
[00396] Methods of preparing polynucleotides containing one or more modified phosphorus- containing intemucleoside linkage are known in the art. See, e.g., U.S. Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,195; 5,188,897; 5,264,423; 5,276,019; 5,278,302;
5,286,717; 5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925;
5,519,126; 5,536,821; 5,541,316; 5,550,111; 5,563,253; 5,571,799; 5,587,361; 5,625,050;
6,028,188; 6,124,445; 6,160,109; 6,169,170; 6,172,209; 6, 239,265; 6,277,603; 6,326,199; 6,346,614; 6,444,423; 6,531,590; 6,534,639; 6,608,035; 6,683,167; 6,858,715; 6,867,294;
6,878,805; 7,015,315; 7,041,816; 7,273,933; 7,321,029; and U.S. Pat. RE39464, the entire contents of each of which arc hereby incorporated herein by reference for all purposes.
(ii) Modified Non-Phosphorous Containing Internucleoside Linkages
[00397] In some embodiments, the modified intemucleoside linkage does not contain a phosphorous atom. Modified intemucleoside linkages that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl intemucleoside linkages, mixed heteroatoms and alkyl or cycloalkyl intemucleoside linkages, or one or more short chain heteroatomic or heterocyclic intemucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; mcthylcncimino and mcthylcnchydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S, and CH2 component parts.
[00398] Representative non-phosphorous containing intemucleoside linking groups include but are not limited to methylenemethylimino (-CH2-N(CH3)-O-CH2-), thiodiester, thionocarbamate (- O-C(=O)(NH)-S-); siloxane (-O-Sith-O-); and N,N'-dimethylhydrazine (-CH2-N(CH3)-N(CH3)-). [00399] Methods of preparing polynucleotides comprising modified internucleoside linkages do not contain a phosphorous atom are known in the art. See, e.g., U.S. Pat. Nos. 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,64,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, the entire contents of each of which are hereby incorporated herein by reference.
4.3.1.3 Additional Exemplary Nucleotide Modifications
[00400] In some embodiments, the modified agent comprises one or more RNA mimetic in which both the sugar and the internucleoside linkage of the nucleotide units are replaced with novel groups. The nucleobase units are maintained for hybridization with an appropriate nucleic acid target (e.g., a target mRNA). In some embodiments, the RNA mimetic is a peptide nucleic acid (PNA). In PNAs, the ribose moiety of the RNA nucleotide is replaced with an amide containing moiety (e.g., an aminoethylglycine). The nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide. Representative US patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, the entire contents of each of which are hereby incorporated herein by reference. Additional PNA compounds suitable for use in the RNA agents described herein are described in, for example, in Nielsen et al., Science, 1991, 254, 1497-1500, the entire contents of which is incorporated by reference herein for all purposes.
[00401] Potentially stabilizing modifications to the terminal ends of the RNA agents (e.g., described herein) can also be incorporated to agents described herein. For example, N- (acetylaminocaproyl)-4-hydroxyprolinol (Hyp-C6-NHAc), N-(caproyl-4-hydroxyprolinol (Hyp- C6), N-(acetyl-4-hydroxyprolinol (Hyp-NHAc), thymidine-2'-O-deoxythymidine (ether), N- (amlnocaproyl)-4-hydroxyprolinol (Hyp-C6-amino), 2-docosanoyl-uridine-3 "-phosphate, inverted base dT(idT) and others. Such modifications are known in the art. See, e.g., WO2011/005861 , the entire contents of which is incorporated herein by reference.
4.3.2 Extent of Modified Nucleotides
[00402] In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent) are modified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the RNA agent (or any component e.g., nucleic acid molecule) thereof) e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) arc modified. In some embodiments, substantially all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified. In some embodiments, all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified.
[00403] In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense and/or antisense strand are modified. For example, at least 50% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 55% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 60% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 65% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 70% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 75% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 80% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 85% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 90% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 90% of the nucleotides of the sense strand and/or antisense strand may be modified. For example, at least 95% of the nucleotides of the sense strand and/or antisense strand may be modified. In some embodiments, substantially all (or all) of the nucleotides in the sense strand and/or antisense strand are modified. In some embodiments, at least one of the modified nucleotides comprises a modified sugar' (e.g., ribose moiety). In some embodiments, at least one of the modified nucleotides comprises a modified nucleobase. In some embodiments, the sense strand comprises at least one modified internucleoside linkage and/or the antisense strand comprises at least one modified internucleoside linkage.
[00404] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18,
17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
[00405] In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent) are unmodified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified. In some embodiments, substantially all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) arc unmodified. In some embodiments, all of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are unmodified.
[00406] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the RNA agent (or any component (e.g., nucleic acid molecule) thereof) (e.g., described herein, e.g., an antisense strand, a sense strand, a dsRNA agent, RNAi agent, etc.) are modified.
[00407] In some embodiments, the RNA agent (e.g., RNAi agent, antisense strand, sense strand, dsRNA agent (e.g., described herein)) comprises one or more non-naturally internucleoside linkage. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the intemucleoside linkages of the RNA agent (e.g., RNAi agent, antisense strand, sense strand, dsRNA agent (e.g., described herein)) are non-naturally occurring. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the or internucleoside linkages of the RNA agent (e.g., RNAi agent, antisense strand, sense strand, dsRNA agent (e.g., described herein)) are chemically modified.
[00408] In some embodiments, the RNA agent comprises a dsRNA agent comprising a sense strand and an antisense strand. In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand are modified. In some embodiments, substantially all of the nucleotides of the sense strand are modified. In some embodiments, all of the nucleotides of the sense strand arc modified.
[00409] In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified. For example, at least 50% of the nucleotides of the sense strand may be modified. For example, at least 55% of the nucleotides of the sense strand may be modified. For example, at least 60% of the nucleotides of the sense strand may be modified. For example, at least 65% of the nucleotides of the sense strand may be modified. For example, at least 70% of the nucleotides of the sense strand may be modified. For example, at least 75% of the nucleotides of the sense strand may be modified. For example, at least 80% of the nucleotides of the sense strand may be modified. For example, at least 85% of the nucleotides of the sense strand may be modified. For example, at least 90% of the nucleotides of the sense strand may be modified. For example, at least 90% of the nucleotides of the sense strand may be modified. For example, at least 95% of the nucleotides of the sense strand may be modified. In some embodiments, substantially all (or all) of the nucleotides in the sense strand are modified. In some embodiments, at least one of the modified nucleotides comprises a modified sugar (e.g., ribose moiety). In some embodiments, at least one of the modified nucleotides comprises a modified nucleobase. In some embodiments, the sense strand comprises at least one modified intemucleoside linkage and/or the antisense strand comprises at least one modified intemucleoside linkage.
[00410] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of sense strand are unmodified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand are unmodified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the a sense strand are unmodified.
[00411] In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand, are unmodified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are unmodified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand are unmodified. In some embodiments, substantially all of the nucleotides of the sense strand are unmodified. In some embodiments, all of the nucleotides of the sense strand arc unmodified.
[00412] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the sense strand are modified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand are modified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand are modified.
[00413] In some embodiments, the sense strand comprises one or more non-naturally intemucleoside linkage. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the intemucleoside linkages of sense strand are non-naturally occurring. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the or intemucleoside linkages of the sense strand are chemically modified.
[00414] In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are modified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are modified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the antisense strand are modified. In some embodiments, substantially all of the nucleotides of the antisense strand are modified. In some embodiments, all of the nucleotides of the antisense strand are modified.
[00415] In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense and/or antisense strand are modified. For example, at least 50% of the nucleotides of the antisense strand may be modified. For example, at least 55% of the nucleotides of the antisense strand may be modified. For example, at least 60% of the nucleotides of the antisense strand may be modified. For example, at least 65% of the nucleotides of the antisense strand may be modified. For example, at least 70% of the nucleotides of the antisense strand may be modified. For example, at least 75% of the nucleotides of the antisense strand may be modified. For example, at least 80% of the nucleotides of the antisense strand may be modified. For example, at least 85% of the nucleotides of the antisense strand may be modified. For example, at least 90% of the nucleotides of the antisense strand may be modified. For example, at least 90% of the nucleotides of the antisense strand may be modified. For example, at least 95% of the nucleotides of the antisense strand may be modified. In some embodiments, substantially all (or all) of the nucleotides in the antisense strand are modified. In some embodiments, at least one of the modified nucleotides comprises a modified sugar’ (e.g., ribose moiety). In some embodiments, at least one of the modified nucleotides comprises a modified nucleobase. In some embodiments, the sense strand comprises at least one modified intemucleoside linkage and/or the antisense strand comprises at least one modified internucleoside linkage.
[00416] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18,
17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the antisense strand are unmodified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the antisense strand are unmodified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are unmodified.
[00417] In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are unmodified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are unmodified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the antisense strand are unmodified. In some embodiments, substantially all of the nucleotides of the antisense strand are unmodified. In some embodiments, all of the nucleotides of the antisense strand are unmodified. [00418] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the antisense strand are modified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the antisense strand are modified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the antisense strand are modified.
[00419] In some embodiments, antisense strand comprises one or more non-naturally intemucleoside linkage. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the internucleoside linkages of the antisense strand arc non-naturally occurring. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the or internucleoside linkages of the antisense strand are chemically modified.
[00420] In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, substantially all of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, all of the nucleotides of the sense strand and antisense strand are modified.
[00421] In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense and/or sense strand and antisense strand are modified. For example, at least 50% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 55% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 60% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 65% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 70% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 75% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 80% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 85% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 90% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 90% of the nucleotides of the sense strand and antisense strand may be modified. For example, at least 95% of the nucleotides of the sense strand and antisense strand may be modified. In some embodiments, substantially all (or all) of the nucleotides in the sense strand and antisense strand are modified. In some embodiments, at least one of the modified nucleotides comprises a modified sugar (e.g., ribose moiety). In some embodiments, at least one of the modified nucleotides comprises a modified nucleobase. In some embodiments, the sense strand comprises at least one modified intemucleoside linkage and/or the sense strand and antisense strand comprises at least one modified intemucleoside linkage.
[00422] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the sense strand and antisense strand are unmodified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are unmodified.
[00423] In some embodiments, at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, substantially all of the nucleotides of the sense strand and antisense strand are unmodified. In some embodiments, all of the nucleotides of the sense strand and antisense strand are unmodified.
[00424] In some embodiments, not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the of the sense strand and antisense strand are modified. In some embodiments, not more than 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the nucleotides of the sense strand and antisense strand are modified. In some embodiments, not more than 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and antisense strand are modified.
[00425] In some embodiments, sense strand and antisense strand comprises one or more non- naturally intemucleoside linkage. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the internucleoside linkages of the sense strand and antisense strand are non-naturally occurring. In some embodiments, at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the or intemucleoside linkages of the sense strand and antisense strand are chemically modified.
4.4 Conjugates [00426] In some embodiments, the RNA agent {e.g., described herein) {e.g., RNAi agent, dsRNA agent, ssRNA agent, antisense strand, sense strand, antisense oligonucleotide) comprises a heterologous moiety {e.g., operably connected to the RNA agent). Therefore, further provided herein are conjugates comprising an RNA agent {e.g., described herein) {e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) and a heterologous moiety {e.g., operably connected to the RNA agent). It is clear from the disclosure, but for the sake of clarity, the conjugate can comprise a modified agent {e.g., described herein, see, e.g., § 4.3).
[00427] In some embodiments, the heterologous moiety modifies one or more properly of the RNA agent {e.g., described herein) {e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) to which it is conjugated. Exemplary properties include, but are not limited to, pharmacodynamics, pharmacokinetics, stability, absorption, activity, tissue distribution, cellular distribution, cellular uptake, charge, half-life, clearance, and binding affinity to a target nucleic acid molecule {e.g., a target mRNA).
[00428] In some embodiments, the heterologous moiety enhances the distribution and/or uptake {e.g., into a cell, e.g., into a cell in a subject) of the RNA agent {e.g., described herein) {e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) {e.g., as compared to an RNA agent that lacks the heterologous moiety). In some embodiments, the heterologous moiety alters {e.g., extends) the lifetime {e.g., in vivo) of the RNA agent {e.g., described herein) {e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) {e.g., as compared to an RNA agent that lacks the heterologous moiety). In some embodiments, the heterologous moiety provides an enhanced affinity for a selected target, e.g., a selected molecule, cell type, compartment {e.g., cell type, tissue, organ or region of the body) {e.g., as compared to an RNA agent that lacks the heterologous moiety).
[00429] In some embodiments, the heterologous moiety enhances the activity {e.g., in a cell, e.g., in a cell in a subject) of the RNA agent {e.g., described herein) {e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) {e.g., as compared to an RNA agent that lacks the heterologous moiety). Activity can include, e.g., degradation of a target mRNA {e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 mRNA), inhibition of expression of a target gene {e.g., a BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6 gene), and/or reduction in the expression of a target gene {e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 gene).
[00430] In some embodiments, the heterologous moiety imparts a new property on the RNA agent {e.g., described herein) {e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) to which it is conjugated. For example, fluorophores or reporter groups that enable detection of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
[00431] It is to be understood the heterologous moieties can impart multiple (e.g., any combination of the foregoing) properties of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
[00432] In some embodiments, wherein the RNA agent is a dsRNA agent comprising a double stranded region, the heterologous moiety does not take part in, does not alter, and/or does not interfere with, the creation of a double strand region.
4.4.1 Heterologous Moieties
[00433] Heterologous moieties, include for example, but are not limited to, carbohydrates, peptides, proteins (e.g., antibodies or functional fragments or variants thereof; ligands (e.g., of a target receptor)), lipids, polymers, small molecules, intcrcalators, reporter molecules, polyamincs, polyamides, vitamin moieties, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipophilic groups, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores, and dyes.
[00434] In some embodiments, the heterologous moiety is a carbohydrate, peptide, protein (e.g., antibody or functional fragment or variant thereof, e.g., ligand (e.g., of a target receptor)), lipid, polymer, small molecule, or any combination thereof. In some embodiments, the heterologous moiety comprises an active drug substance. In some embodiments, the heterologous moiety does not contain an active drug substance.
4.4.1.1 T ar eting Moieties
[00435] In some embodiments, the heterologous moiety is a targeting moiety. In some embodiments, the targeting moiety enhances distribution of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) to a target cell (or population of cells), tissue, and/or organ (e.g., as compared to an RNA agent that lacks the targeting moiety). In some embodiments, the targeting moiety enhances the uptake of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) into a target cell (or population of cells) (e.g., as compared to an RNA agent that lacks the targeting moiety). In some embodiments, the targeting moiety provides enhanced affinity for a selected target, e.g., molecule, cell, cell type, compartment, e.g., a cellular or organ compartment, tissue, organ or region of the body, (e.g., as compared to an RNA agent that lacks the targeting moiety).
[00436] In some embodiments, the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed on the surface of a target cell, tissue, and/or organ. In some embodiments, the target molecule is a protein, carbohydrate, or lipid. In some embodiments, the target molecule is a receptor.
(i) Immune Cell Targeting Moieties
[00437] In some embodiments, the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by one or more population of immune cells (e.g., on the surface of the surface of one or more population of immune cells). In some embodiments, the targeting moiety specifically binds to a target molecule protein (e.g., receptor) expressed on the surface of one or more population of immune cells.
[00438] In some embodiments, the immune cells are within the microenvironment of a tumor. In some embodiments, the immune cells are characterized as tumor infiltrating lymphocytes.
[00439] In some embodiments, the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by T-cells (e.g., on the surface of the surface of T- cells). In some embodiments, the targeting moiety specifically binds to a target molecule protein (e.g., receptor) expressed on the surface of T-cells. In some embodiments, the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by CD4+ T-cells (e.g., on the surface of the surface of CD4+ T-cells). In some embodiments, the targeting moiety specifically binds to a target molecule protein (e.g., receptor) expressed on the surface of CD4+ T-cells. In some embodiments, the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by CD8+ T-cells (e.g., on the surface of the surface of CD8+ T-cells). In some embodiments, the targeting moiety specifically binds to a target molecule protein (e.g., receptor) expressed on the surface of CD8+ T-cells.
[00440] In some embodiments, the T-cells are within the microenvironment of a tumor. In some embodiments, the T-cells are characterized as tumor infiltrating T-cells. In some embodiments, the CD4+ T-cells are within the microenvironment of a tumor. In some embodiments, the CD4+ T- cells are characterized as tumor infiltrating CD4+ T-cells. In some embodiments, the CD8+ T- cells are within the microenvironment of a tumor. In some embodiments, the CD8+ T-cells are characterized as tumor infiltrating CD8+ T-cells.
[00441] In some embodiments, the targeting moiety specifically binds to a target molecule (e.g., protein, carbohydrate, lipid, etc.) expressed by B-cells (e.g., on the surface of the surface of B- cells). In some embodiments, the targeting moiety specifically binds to a target molecule protein (e.g., receptor) expressed on the surface of B-cells. In some embodiments, the B-cells are within the microenvironment of a tumor. In some embodiments, the B-cells are characterized as tumor infiltrating B-cells.
4.4.2 Linkers
[00442] In some embodiments, the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) is directly attached to the heterologous moiety (e.g., targeting moiety) (e.g., directly attached through a single chemical bond). In some embodiments, the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) is indirectly attached to the heterologous moiety (e.g., targeting moiety). In some embodiments, the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) is indirectly attached to the heterologous moiety via a linker.
[00443] Suitable linkers for use in the conjugates described herein are known in the art and can be evaluated by a person of ordinary skill in the ait using standard methods. Exemplary linkers and components thereof for use in the conjugates described herein are also described below.
[00444] Linkers typically comprise a direct bond or an atom such as oxygen or sulfur, a unit such as NR8, C(O), C(O)NH, SO, SO2, SO2NH or a chain of atoms, such as, but not limited to, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl, alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl, alkenylheteroarylalkenyl, alkenylheteroarylalkynyl, alkynylheteroarylalkyl. alkynylheteroarylalkenyl, alkynylheteroarylalkynyl, alkylheterocyclylalkyl alkylheterocyclylalkenyl, alkylhererocyclylalkynyl, alkenylheterocyclylalkyl. alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl. alkynylheterocyclyl alkenyl, alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl, alkylhctcroaryl, alkcnylhctcroaryl, alkynylhcrcroaryl, which one or more methylenes can be inteiTupted or terminated by O, S, S(O), SO2, N(R8), C(O), substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocyclic; where R8 is hydrogen, acyl, aliphatic, or substituted aliphatic. In one embodiment, the linker is about 1-24 atoms, 2-24, 3-24, 4-24, 5-24, 6-24, 6-18, 7-18, 8-18, 7-17, 8-17, 6-16, 7-17, or 8-16 atoms.
[00445] In some embodiments, the linker comprises ethylene glycol, nucleosides, or amino acid units. In some embodiments, the linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In some embodiments, the linker comprises groups selected from alkyl, amino, oxo, amide and ether groups. In some embodiments, the linker comprises groups selected from alkyl and amide groups. In some embodiments, the linker comprises groups selected from alkyl and ether groups. In some embodiments, the linker comprises at least one phosphorus moiety. In some embodiments, the linker comprises at least one phosphate group. In some embodiments, the linker comprises at least one neutral linking group. Exemplary linkers include but are not limited to pyrrolidine, 8-amino- 3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane- 1 -carboxylate (SMCC), 6-aminohexanoic acid (AHEX or AHA). Additional exemplary linkers include but are not limited to substituted or unsubstituted Ci-Cw alkyl, substituted or unsubstituted C2-C10 alkenyl or substituted or unsubstituted C2-C10 alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.
[00446] In some embodiments, the linker is bifunctional. In general, a bifunctional linker comprises at least two functional groups. One of the functional groups is selected to react with a particular' site on an RNA agent (e.g., described herein) and the other is selected to react with a heterologous moiety (e.g., described herein). Examples of functional groups used in a bifunctional linkers include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In some embodiments, bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.
[00447] In some embodiments, the linker is a monovalent linker, a bivalent linker, a trivalent linker, or a tetravalent linker. 4.4.2.1 Cleavable Linkers
[00448] In some embodiments, the linker is non-cleavable. In some embodiments, the linker is cleavable. Cleavable linkers contain at least one (or a plurality of) cleavable bonds that are susceptible to one or more cleavage agent. Exemplary classes of cleavable linkers include, but are not limited to, redox clcavablc linkers, phosphate based clcavablc linkers, acid clcavablc linkers, ester-based cleavable linkers, and peptide-based cleavable linkers. In certain embodiments, a cleavable bond is selected from among: an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, or a disulfide.
[00449] Cleavable linkers may be advantageous when a stable conjugate is desired under a first set of conditions but under a second set of conditions it is advantageous to release the RNA agent (e.g., described herein) from the heterologous moiety (e.g., described herein). For example, in some embodiments, it may be desirable to have a sufficiently stable conjugate outside of a cell (e.g., within a subject (e.g., within the blood or serum of a subject)), and upon entry into a cell (e.g., a target cell (e.g., a target cell within a subject)) have the linker cleaved to release the RNA agent (e.g., described herein) from the heterologous moiety (e.g., described herein). In some embodiments, the linker is not cleaved (or is cleaved at a lower rate) under a first condition relative to under a second condition. In some embodiments, the first condition is within the blood (e.g., of a subject) (or in an in vitro system sufficient to mimic the conditions of the blood within a subject) and the second condition is with a cell (e.g., a cell within a subject) (or in an in vitro system sufficient to mimic the conditions of a cell within a subject).
[00450] The suitability of a cleavable linker can be assessed by standard methods known in the art. In general, the suitability of a cleavable linker can be evaluated by testing the ability of a cleavage agent (or condition) to cleave the candidate linker (e.g., the cleavage bond(s)). In some embodiments, it may be desirable to further test the ability of the linker to resist cleavage under a certain condition (e.g., within the blood or serum of subject, when in contact with a non-target cell, tissue, organ).
[00451] In some embodiments, the linker is a redox cleavable linker that is cleaved upon reduction or oxidation. An example of a reductively cleavable linker is a disulphide (-S-S-) containing linker. Redox cleavable linkers can be evaluated using methods analogous to those described above.
[00452] In some embodiments, the linker is a phosphate-based cleavable linker. A phosphate- based cleavable linker is cleaved by agents that degrade or hydrolyze the phosphate group. For example, in cells, enzymes such as phosphatases arc capable of cleaving phosphate groups. Examples of phosphate -based linkers include those comprising any of the following -O- P(O)(ORk)-O-, -OP(S)(ORk)-O-, -O-P(S)(SRk)-O-, -S-P(O)(ORk)-O-, -O-P(O)(ORk)-S-, -S- P(O)(ORk)-S-, -OP(S)(ORk)-S-, -S-P(S)(ORk)-O-, -O-P(O)(Rk)-O-, -O-P(S)(Rk)-O-, -S- P(O)(Rk)-O-, -S-P(S)(Rk)-O-, -S-P(O)(Rk)-S-, -O-P(S)( Rk)-S-, wherein Rk at each occurrence can be, independently, C1-C20 alkyl, C1-C20 haloalkyl, C6-C10 aryl, or C7-C12 aralkyl. Exemplary embodiments include are -OP(O)(OH)-O-, -O-P(S)(OH)-O-, -O-P(S)(SH)-O-, -S- P(O)(OH)-O-, -O-P(O)(OH)-S-, -S-P(O)(OH)-S-, -O-P(S)(OH)-S-, -S-P(S)(OH)-O-, -O-P(O)(H)- O-, -O-P(S)(H)-O-, -S-P(O)(H)-O, -S-P(S)(H)-O-, -S-P(O)(H)-S-, or -O-P(S)(H)-S-. Phosphate based cleavable linker can be evaluated using methods analogous to those described above.
[00453] In some embodiments, the linker is an acid cleavable linker. An acid cleavable linker is cleaved under acidic conditions. For example, in some embodiments the acid cleavable linker can be cleaved in an acidic environment with a pH of about 6.5 or less (e.g., about 6.0, 5.5, 5.0, or less). In some embodiments the acid cleavable linker can be cleaved by enzymes that can act as a general acid. In a cell (e.g., within a subject), specific low pH organelles, such as endosomes and lysosomes can provide a cleaving environment for acid cleavable linkers. Examples of acid cleavable linkers include but are not limited to hydrazones, esters, and esters of amino acids. Acid cleavable groups can have the general formula -C=NN-, C(O)O, or -OC(O). Acid cleavable linkers can be evaluated using methods analogous to those described above.
[00454] In some embodiments, the linker is an ester-based cleavable linker. An ester-based cleavable linker is cleaved by enzymes such as esterases and amidases in cells. Examples of ester- based cleavable include, but are not limited to, esters of alkylene, alkenylene and alkynylene groups. The cleavable bonds of ester cleavable linkers have the general formula -C(O)O- or - OC(O)-. Ester-based cleavable linkers can be evaluated using methods analogous to those described above.
[00455] In some embodiments, the linker is a peptide-based cleavable linker. A peptide-based cleavable linker is cleaved by enzymes such as peptidases and proteases (e.g., present in cells (e.g., cells within a subject)). Peptide-based cleavable linkers comprise peptide bonds formed between amino acids to yield polypeptides (e.g., dipeptides, tripeptides, etc.). As known in the art, peptide bonds. The peptide bonds (i.e., the amide bond) of the peptide linker is generally the site of cleavage. Peptide-based cleavable linkers can be evaluated using methods analogous to those described above.
4.4.3 Orientation
[00456] The heterologous moiety may be attached at any suitable position to the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
[00457] In some embodiments, the heterologous moiety is conjugated to the 5' end of the RNA agent (e.g., described herein) e.g., RNAi agent, dsRNA agent, antisense strand, sense strand). In some embodiments, the heterologous moiety is conjugated to the 3' end of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand). In some embodiments, a first heterologous moiety is conjugated to the 5' end of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) and a second heterologous moiety is conjugated to the 3' end of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand). The first and second heterologous moieties can be the same or different. In some embodiments, the heterologous moiety is conjugated to an internal site of the RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
[00458] In some embodiments, the RNA agent (e.g., RNAi agent, dsRNA agent) comprises an antisense strand. In some embodiments, the heterologous moiety is conjugated to the 5' end of the antisense strand. In some embodiments, the heterologous moiety is conjugated to the 3' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to the 5' end of the antisense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand. The first and second heterologous moieties can be the same or different. In some embodiments, the heterologous moiety is conjugated to an internal site of the antisense strand.
[00459] The heterologous moiety may be attached to the 3' end of the sense and/or antisense strand. The heterologous moiety may be attached to the 5' end of the sense and/or antisense strand. The heterologous moiety may be attached to at an internal site of the sense and/or antisense strand. The heterologous moiety may be attached to the 3' end of the sense and antisense strand. The heterologous moiety may be attached to the 5' end of the sense and antisense strand. The heterologous moiety may be attached to at an internal site of the sense and antisense strand.
[00460] In some embodiments, the RNA agent (e.g., RNAi agent, dsRNA agent) comprises a sense strand. Tn some embodiments, the heterologous moiety is conjugated to the 5' end of the sense strand. In some embodiments, the heterologous moiety is conjugated to the 3' end of the sense strand. In some embodiments, a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the sense strand. The first and second heterologous moieties can be the same or different. In some embodiments, the heterologous moiety is conjugated to an internal site of the sense strand.
[00461] In some embodiments, the RNA agent (e.g., RNAi agent) comprises a dsRNA agent comprising a sense strand and an antisense strand, hi some embodiments, the heterologous moiety is conjugated to the 5' end of the sense strand. In some embodiments, the heterologous moiety is conjugated to the 3' end of the sense strand. In some embodiments, a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the sense strand. The first and second heterologous moieties can be the same or different. In some embodiments, the heterologous moiety is conjugated to an internal site of the sense strand. In some embodiments, the heterologous moiety is conjugated to the 5' end of the sense strand. In some embodiments, the heterologous moiety is conjugated to the 3' end of the sense strand. In some embodiments, a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the sense strand. The first and second heterologous moieties can be the same or different. In some embodiments, the heterologous moiety is conjugated to an internal site of the sense strand.
[00462] In some embodiments, a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to the 3' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to the 5' end of the sense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to the 3' end of the sense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand. The first and second heterologous moieties can be the same or different.
[00463] In some embodiments, a first heterologous moiety is conjugated to an internal site of the sense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to an internal site of the sense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to an internal site of the antisense strand and a second heterologous moiety is conjugated to the 3' end of the antisense strand. In some embodiments, a first heterologous moiety is conjugated to an internal site of the antisense strand and a second heterologous moiety is conjugated to the 5' end of the antisense strand. The first and second heterologous moieties can be the same or different.
4.5 Activity of RNA Agents & Conjugates Thereof
[00464] In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.). In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 90%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 50%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 75%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 80%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 90%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) inhibits expression of a target gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject (e.g., a mammalian subject, e.g., a primate, human, non-human primate, mouse, rat, etc.) by at least about 95%. In some embodiments, the RNA agent (e.g., RNAi agent, e.g., dsRNA agent) mediates degradation of a target mRNA (e.g., a BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 (e.g., hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6) mRNA). [00465] Any one or more of the above activities can be evaluated in vitro, ex vivo, or in vivo. Any one or more of the above activities can be evaluated by standard methods known in the art. For example, by PCR (e.g., qPCR), branched DNA assays, or by a protein-based methods (such as immunofluorescence analysis (using, e.g., western blotting or flow cytometric techniques). In some embodiments, inhibition of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6 (e.g., hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6)) expression is determined by qPCR.
[00466] In some embodiments, the cell is an immune cell. Exemplary immune cells include, T cells (e.g., CD4+ T cells, CD8+ T cells), B-cells, tumor infiltrating lymphocytes (e.g., tumor infiltrating T cells (e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells)). In some embodiments, the cell is a T-cell. In some embodiments, the cell is a tumor infiltrating lymphocyte. In some embodiments, the cell is a tumor infiltrating T cell. In some embodiments, the cell is a tumor infiltrating CD4+ T cell. In some embodiments, the cell is a CD8+ tumor infiltrating T cell. In some embodiments, the cell is B-cell.
4.6 Methods of Making RNA Agents & Conjugates Thereof [00467] An RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand) can be synthesized by standard methods known in the ait (e.g., chemical synthesis (e.g., solid phase synthesis)). See, e.g., “Current protocols in nucleic acid chemistry,” Beaucage, S. L. et al. (Edrs.), John Wiley & Sons, Inc., New York, N.Y., USA, and See, e.g., Dong Y, Siegwart DJ, Anderson DG. Strategies, design, and chemistry in siRNA delivery systems. Adv Drug Deliv Rev. 2019 Apr;144:133-147. doi: 10.1016/j.addr.2019.05.004. Epub 2019 May 15. PMID: 31102606; PMCID: PMC6745264, the entire contents of each of which is incorporated by reference herein for all purposes. As such, further provided herein are methods of making an RNA agent described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand).
[00468] For example, single stranded nucleic acid molecules (e.g., described herein) (e.g., antisense strands, sense strands) can be prepared using solution-phase or solid-phase organic synthesis or both. dsRNA agents (e.g., described herein) can be prepared using a two-step procedure, wherein the individual strands of the dsRNA agent are prepared separately and subsequently annealed. The individual strands of the dsRNA agent can be prepared using solutionphase or solid-phase organic synthesis or both. Regardless of the method of synthesis, the RNA agents (e.g., described herein) (e.g., dsRNA agents described herein) can be prepared in a solution (e.g., an aqueous or organic solution) that is appropriate for formulation. For example, the dsRNA agent can be precipitated and redissolved in pure double-distilled water, and lyophilized. The lyophilized dsRNA agent can be resuspended in a solution appropriate for the intended formulation process.
[00469] Likewise, conjugates (e.g., described herein) can be synthesized utilizing standard methods known in the ait. See, e.g., Dong Y, Siegwart DJ, Anderson DG. Strategies, design, and chemistry in siRNA delivery systems. Adv Drug Deliv Rev. 2019 Apr;144: 133-147. doi: 10.1016/j.addr.2019.05.004. Epub 2019 May 15. PMID: 31102606; PMCID: PMC6745264, the entire contents of which is incorporated herein by reference for all purposes. A person of ordinary skill in the art can determine the appropriate conjugation method based on e.g., the heterologous moiety and the RNA agent to be conjugated. For example, standard conjugation methods include, e.g., parallel synthesis methods and linear synthesis methods.
4.7 Vectors
[00470] In some embodiments, one or more of the RNA agents described herein (e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands) (see, e.g., §§ 4.2, 4.2.2) arc contained in a vector e.g., a non-viral vector (e.g., a plasmid), a viral vector). Thus, in one aspect, also provided herein are vectors (e.g., non-viral vectors (e.g., plasmids) viral vectors) comprising one or more agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand) (see, e.g., §§ 4.2, 4.3). Such vectors can be easily manipulated by methods well known to the ordinary person of skill in the art. The vector used can be any vector that is suitable for cloning nucleic acid molecules that can be used for transcription of the nucleic acid molecule of interest (e.g., an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand) (see, e.g., §§ 4.2, 4.3).
[00471] In some embodiments, the vector is a viral vector. Viral vectors include both RNA and DNA based vectors. The vectors can be designed to meet a variety of specifications. For example, viral vectors can be engineered to be capable or incapable of replication in prokaryotic and/or eukaryotic cells. In some embodiments, the vector is replication deficient. In some embodiments, the vector is replication competent. Vectors can be engineered or selected that either will (or will not) integrate in whole or in part into the genome of host cells, resulting (or not (e.g., episomal expression)) in stable host cells comprising the desired nucleic acid in their genome.
[00472] Exemplary viral vectors include, but are not limited to, adenovirus vectors, adeno- associated virus vectors, lentivirus vectors, retrovirus vectors, poxvirus vectors, parapoxivirus vectors, vaccinia virus vectors, fowlpox virus vectors, herpes virus vectors, adeno-associated virus vectors, alphavirus vectors, lentivirus vectors, rhabdovirus vectors, measles virus, Newcastle disease virus vectors, picornaviruses vectors, anellovectors, or lymphocytic choriomeningitis virus vectors. In some embodiments, the viral vector is an adenovirus vector, adeno-associated virus vector, lentivirus vector, anellovector (as described, for example, in U.S. Pat. No. 11,446,344, the entire contents of which is incorporated by reference herein for all purposes).
[00473] In some embodiments, the vector is an adenoviral vector (e.g., human adenoviral vector, e.g., HAdV or AdHu). In some embodiments, the adenovirus vector has the El region deleted, rendering it replication-deficient in human cells. Other regions of the adenovirus such as E3 and E4 may also be deleted. Exemplary adenovirus vectors include, but are not limited to, those described in e.g., W02005071093 or WQ2006048215, the entire contents of each of which is incorporated by reference herein for all purposes. In some embodiments, the adenovirus-based vector used is a simian adenovirus, thereby avoiding dampening of the immune response after vaccination by pre-existing antibodies to common human entities such as AdHu5. Exemplary, simian adenovirus vectors include AdCh63 (see, e.g., W02005071093, the entire contents of which is incorporated by reference herein for all purposes) or AdCh68.
[00474] Viral vectors can be generated through the use of a packaging/producer cell line (e.g., a mammalian cell line) using standard methods known to the person of ordinary skill in the art. Generally, a nucleic acid construct (e.g., a plasmid) encoding the transgene (e.g., an RNA agent described herein) (along with additional elements e.g., a promoter, inverted terminal repeats (ITRs) flanking the transgene, a plasmid encoding e.g., viral replication and structural proteins, along with one or more helper plasmids a host cell (e.g., a host cell line) are transfected into a host cell line (i.e., the packing/producer cell line). In some instances, depending on the viral vector, a helper plasmid may also be needed that include helper genes from another virus (e.g., in the instance of adeno-associated viral vectors). Eukaryotic expression plasmids are commercially available from a variety of suppliers, for example the plasmid series: pcDNA™, pCR3.1 ™, pCMV™, pFRT™, pVAXl ™, pCI™, Nanoplasmid™, and Pcaggs. The person of ordinary skill in the art is aware of numerous transfection methods and any suitable method of transfection may be employed (e.g., using a biochemical substance as carrier (e.g., lipofectamine), by mechanical means, or by electroporation,). The cells are cultured under conditions suitable and for a sufficient time for plasmid expression. The viral particles may be purified from the cell culture medium using standard methods known to the person of ordinary skill in the art. For example, by centrifugation followed by e.g., chromatography or ultrafiltration.
[00475] In some embodiments, the vector is a plasmid. A person of ordinary skill in the art is aware of suitable plasmids for expression of the DNA of interest. For example, Suitable plasmid DNA may be generated to allow efficient production of the encoded RNA in cell lines, e.g., in insect cell lines, for example using vectors as described in W02009150222A2 and as defined in PCT claims 1 to 33, the disclosure relating to claim 1 to 33 of W02009150222A2 the entire contents of which is incorporated by reference herein for all purposes.
4.8 Cells
[00476] Provided herein is a cell (e.g., host cell) or population of cells (e.g., a population of host cells) comprising any one more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand) (see, e.g., §§ 4.2, 4.3); a vector described herein (see, e.g., § 4.7); a conjugate described herein (see, e.g., § 4.4); a carrier described herein (see, e.g., § 4.9); a pharmaceutical composition described herein (see, e.g., § 4.10), and/or a cell described herein (see, e.g., § 4.8); or any combination thereof. In some embodiments, the cell or population of cells is in vitro. In some embodiments, the cell or population of cells is in vivo. In some embodiments, the cell or population of cells is ex vivo. In some embodiments, the cell or population of cells are derived from a subject (e.g., a human subject. In some embodiments, the subject is a human, mouse, rat, primate, or non-human primate.
[00477] In some embodiments, the cell (or population of cells) is an immune cell. In some embodiments, the cell (or population of cells) is a subset of immune cells.
[00478] In some embodiments, the immune cells are within the microenvironment of a tumor. In some embodiments, the immune cells are characterized as tumor infiltrating lymphocytes.
[00479] In some embodiments, the cell (or population of cells) is a CD4+ T cell. In some embodiments, the cell (or population of cells) is a CD8+ T cell. In some embodiments, the population of cells comprises is CD8+ T cells and CD4+ T cells.
[00480] In some embodiments, the T-cells are within the microenvironment of a tumor. In some embodiments, the T-cells are characterized as tumor infiltrating T-cells. In some embodiments, the CD4+ T-cells are within the microenvironment of a tumor. In some embodiments, the CD4+ T- cells are characterized as tumor infiltrating CD4+ T-cells. In some embodiments, the CD8+ T- cells are within the microenvironment of a tumor. In some embodiments, the CD8+ T-cells are characterized as tumor infiltrating CD8+ T-cells.
[00481] In some embodiments, the cell (or population of cells) is a B cell. In some embodiments, the cell (or population of cells) is a B cell. In some embodiments, the population of cells comprises is B cells.
[00482] In some embodiments, the B-cells are within the microenvironment of a tumor. In some embodiments, the B-cells are characterized as tumor infiltrating B-cells.
4.9 Carriers
[00483] In some embodiments, one or more of the RNA agents described herein (e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands (or a conjugate comprising the same)) or a vector comprising any of the foregoing is formulated within one or more carrier. [00484] Therefore, further provided herein are carriers comprising any one or more of the RNA agents described herein (e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands (or a conjugate comprising the same)) or a vector comprising any of the foregoing.
[00485] Any of the foregoing (e.g., one or more of the RNA agents described herein (e.g., RNAi agents, double stranded RNA (dsRNA) agents, sense strands, antisense strands (or a conjugate comprising the same)) or a vector comprising any of the foregoing) can be encapsulated within a carrier, chemically conjugated to a carrier, associated with the carrier. In this context, the term “associated” refers to the essentially stable combination of an RNA agent described herein (or a conjugate comprising the same) (or a vector comprising the same) with one or more molecules of a carrier (e.g., one or more lipids of a lipid-based carrier, e.g., an LNP, liposome, lipoplex, and/or nanoliposome) into larger complexes or assemblies without covalent binding. In this context, the term “encapsulation” refers to the incorporation of an RNA agent described herein (or a conjugate comprising the same) (or a vector comprising the same) into a carrier (e.g., a lipid-based carrier, e.g., an LNP, liposome, lipoplex, and/or nanoliposome) wherein the RNA agent described herein (or the conjugate comprising the same) (or the vector comprising the same) is entirely contained within the interior space of the carrier (e.g., the lipid-based carrier, e.g., the LNP, liposome, lipoplex, and/or nanoliposome).
[00486] Exemplary carriers includes, but are not limited to, lipid-based carriers (e.g., lipid nanoparticles (LNPs), liposomes, lipoplexes, and nanoliposomes). In some embodiments, the carrier is a lipid-based carrier. In some embodiments, the carrier is an LNP. In some embodiments, the LNP comprises a cationic lipid, a neutral lipid, a cholesterol, and/or a PEG lipid. Lipid based carriers are further described below in § 4.9.1.
4.9.1 Lipid Based Carriers/Lipid Nanoformulations
[00487] In some embodiments, an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) is encapsulated or associated with one or more lipids (e.g., cationic lipids and/or neutral lipids), thereby forming lipid-based carriers such as lipid nanoparticles (LNPs), liposomes, lipoplexes, or nanoliposomes.
[00488] In some embodiments, an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) is encapsulated in one or more lipids (e.g., cationic lipids and/or neutral lipids), thereby forming lipid-based earners such as lipid nanoparticles (LNPs), liposomes, lipoplexes, or nanoliposomes. In some embodiments, an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) is associated with one or more lipids (e.g., cationic lipids and/or neutral lipids), thereby forming lipid-based carriers such as lipid nanoparticles (LNPs), liposomes, lipoplexes, or nanoliposomes. In some embodiments, an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) is encapsulated in LNPs (e.g., as described herein).
[00489] The RNA agents (e.g., RNAi agents, dsRNA agents, sense strands, antisense strands (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) described herein may be completely or partially located in the interior space of the LNPs, liposomes, lipoplexes, and/or nanoliposomes, within the lipid layer/membrane, or associated with the exterior surface of the lipid layer/membrane. One purpose of incorporating an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) into LNPs, liposomes, lipoplexes, and/or nanoliposomes is to protect the RNA agent from an environment which may contain enzymes or chemicals or conditions that degrade the RNA agent from molecules or conditions that cause the rapid excretion of the RNA agent. Moreover, incorporating an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) into LNPs, liposomes, lipoplexes, and/or nanoliposomes may promote the uptake of the RNA agent, and hence, may enhance the therapeutic effect of the RNA agent (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing). Accordingly, incorporating an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing), into LNPs, liposomes, lipoplexes, and/or nanoliposomes may be particularly suitable for a pharmaceutical composition described herein, e.g., for intramuscular and/or intradermal administration. [00490] In some embodiments, an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) is formulated into a lipid-based carrier (or lipid nanoformulation). In some embodiments, the lipid-based carrier (or lipid nanoformulation) is a liposome or a lipid nanoparticle (LNP). In one embodiment, the lipid-based carrier is an LNP.
[00491] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises a cationic lipid (e.g., an ionizable lipid), a non-cationic lipid (e.g., phospholipid), a structural lipid (e.g., cholesterol), and a PEG-modified lipid. In some embodiments, the lipid-based carrier (or lipid nanoformulation) contains one or more agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing), or a pharmaceutically acceptable salt thereof.
[00492] As described herein, suitable compounds to be used in the lipid-based carrier (or lipid nanoformulation) include all the isomers and isotopes of the compounds described above, as well as all the pharmaceutically acceptable salts, solvates, or hydrates thereof, and all crystal forms, crystal form mixtures, and anhydrides or hydrates.
[00493] In addition to one or more agent described herein, the lipid-based carrier (or lipid nanoformulation) may further include a second lipid. In some embodiments, the second lipid is a cationic lipid, a non-cationic (e.g., neutral, anionic, or zwitterionic) lipid, or an ionizable lipid.
[00494] One or more naturally occurring and/or synthetic lipid compounds may be used in the preparation of the lipid-based carrier (or lipid nanoformulation).
[00495] The lipid-based earner (or lipid nanoformulation) may contain positively charged (cationic) lipids, neutral lipids, negatively charged (anionic) lipids, or a combination thereof.
4.9.1.1 Cationic Lipids (Positively Charged) and Ionizable Lipids
[00496] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises one or more cationic lipids, e.g., a cationic lipid that can exist in a positively charged or neutral form depending on pH, or an amine-containing lipid that can be readily protonated. In some embodiments, the cationic lipid is a lipid capable of being positively charged, e.g., under physiological conditions.
[00497] Exemplary cationic lipids include one or more amine group(s) which bear the positive charge. Examples of positively charged (cationic) lipids include, but are not limited to, N,N'- dimethyl-N,N'-dioctacyl ammonium bromide (DDAB) and chloride DDAC), N-(l-(2,3- dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), 3P-[N-(N',N'- dimcthylaminocthyl)carbamoyl) cholesterol (DC-chol), l,2-diolcoyloxy-3-[trimcthylammonio]- propane (DOTAP), l,2-dioctadecyloxy-3-[trimethylammonio] -propane (DSTAP), and 1,2- dioleoyloxypropyl-3-dimethyl-hydroxy ethyl ammonium chloride (DORI), N,N-dioleyl-N,N- dimethylammonium chloride (DODAC), N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1 ,2-Dioleoyl-3-Dimethylammonium-propane (DODAP), 1 ,2-Dioleoylcarbamyl-3-
Dimethylammonium-propane (DOCDAP), 1 ,2-Dilineoyl-3-Dimethylammonium-propane (DLINDAP), 3-Dimethylamino-2-(Cholest-5-en-3-beta-oxybutan-4-oxy)-l-(cis,cis-9,12- octadecadienoxy)propane (CLinDMA), 2-[5'-(cholest-5-en-3-beta-oxy)-3'-oxapentoxy)-3- dimethyl-l-(cis, cis-9',12'-octadecadienoxy)propane (CpLin DMA), N,N-Dimethyl-3,4- dioleyloxybenzylamine (DMOBA), and the cationic lipids described in e.g. Martin et al., Current Pharmaceutical Design, pages 1-394, the entire contents of which are incorporated by reference herein for all purposes. In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises more than one cationic lipid.
[00498] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises a cationic lipid having an effective pKa over 6.0. In some embodiments, the lipid-based carrier (or lipid nanoformulation) further comprises a second cationic lipid having a different effective pKa (e.g., greater than the first effective pKa) than the first cationic lipid.
[00499] In some embodiments, cationic lipids that can be used in the lipid-based carrier (or lipid nanoformulation) include, for example those described in Table 4 of WO 2019/217941, the entire contents of which are incorporated by reference herein for all purposes.
[00500] In some embodiments, the cationic lipid is an ionizable lipid (e.g., a lipid that is protonated at low pH, but that remains neutral at physiological pH). In some embodiments, the lipid-based carrier (or lipid nanoformulation) may comprise one or more additional ionizable lipids, different than the ionizable lipids described herein. Exemplary ionizable lipids include, but are not limited to,
(see WO2017004143A1, the entire contents of which is incorporated herein by reference for all purposes).
[00501] In some embodiments, the lipid-based carrier (or lipid nanoformulation) further comprises one or more compounds described by WO 2021/113777 (e.g., a lipid of Formula (3) such as a lipid of Table 3 of WO 2021/113777), the entire contents of which are incorporated by reference herein for all purposes.
[00502] In one embodiment, the ionizable lipid is a lipid disclosed in Hou, X., et al. Nat Rev Mater 6, 1078-1094 (2021). https://doi.org/10.1038/s41578-021-00358-0 e.g., L319, C12-200, and DLin-MC3-DMA), (the entire contents of which are incorporated by reference herein for all purposes).
[00503] Examples of other ionizable lipids that can be used in lipid-based carrier (or lipid nanoformulation) include, without limitation, one or more of the following formulas: X of US 2016/0311759; I of US 20150376115 or in US 2016/0376224; Compound 5 or Compound 6 in US 2016/0376224; T, TA, or II of US 9,867,888; I, II or III of US 2016/0151284; I, IA, II, or IIA of US 2017/0210967; I-c of US 2015/0140070; A of US 2013/0178541; I of US 2013/0303587 or US 2013/0123338; I of US 2015/0141678; II, III, IV, or V of US 2015/0239926; I of US 2017/0119904; I or II of WO 2017/117528; A of US 2012/0149894; A of US 2015/0057373; A of WO 2013/116126; A of US 2013/0090372; A of US 2013/0274523; A of US 2013/0274504; A of US 2013/0053572; A of WO 2013/016058; A of WO 2012/162210; I of US 2008/042973; I, II, III, or IV of US 2012/01287670; I or II of US 2014/0200257; I, II, or III of US 2015/0203446; I or III of US 2015/0005363; I, IA, IB, IC, ID, II, IIA, IIB, IIC, IID, or III-XXIV of US 2014/0308304; of US 2013/0338210; I, II, III, or IV of WO 2009/132131; A of US 2012/01011478; I or XXXV of US 2012/0027796; XIV or XVII of US 2012/0058144; of US 2013/0323269; I of US 2011/0117125; I, II, or III of US 2011/0256175; I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII of US 2012/0202871; I, II, III, IV, V, VI, VII, VIII, X, XII, XIII, XIV, XV, or XVI of US 2011/0076335; I or II of US 2006/008378; I of W02015/074085 (e.g., ATX-002); I of US 2013/0123338; I or X-A-Y-Z of US 2015/0064242; XVI, XVII, or XVIII of US 2013/0022649; I, II, or III of US 2013/0116307; I, II, or III of US 2013/0116307; I or II of US 2010/0062967; I-X of US 2013/0189351; I of US 2014/0039032; V of US 2018/0028664; I of US 2016/0317458; I of US 2013/0195920; 5, 6, or 10 of US 10,221,127; III-3 of WO 2018/081480; 1-5 or 1-8 of WO 2020/081938; I of WO 2015/199952 e.g., compound 6 or 22) and Table 1 therein; 18 or 25 of US 9,867,888; A of US 2019/0136231; II of WO 2020/219876; 1 of US 2012/0027803; OF-02 of US 2019/0240349; 23 of US 10,086,013; cKK-E12/A6 of Miao et al (2020); C12-200 of WO 2010/053572; 7C1 of Dahlman et al (2017); 304-013 or 503-013 of Whitehead et al; TS- P4C2 of U S9, 708, 628; I of WO 2020/106946; I of WO 2020/106946; (1), (2), (3), or (4) of WO 2021/113777; and any one of Tables 1-16 of WO 2021/113777, the entire contents of each of which are incorporated by reference herein for all purposes.
[00504] In some embodiments, the lipid-based carrier (or lipid nanoformulation) further includes biodegradable ionizable lipids, for instance, (9Z,12Z)-3-((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino )propoxy)carbonyl)oxy)methyl)propyl octadeca-9,12-dienoate, also called 3- ((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate). See, e.g., lipids of WO 2019/067992, WO 2017/173054, WO 2015/095340, and WO 2014/136086, the entire contents of each of which are incorporated by reference herein for all purposes. 4.9.1.2 Non-Cationic Lipids (e.g., Phospholipids)
[00505] In some embodiments, the lipid-based carrier (or lipid nanoformulation) further comprises one or more non-cationic lipids. In some embodiments, the non-cationic lipid is a phospholipid. In some embodiments, the non-cationic lipid is a phospholipid substitute or replacement. In some embodiments, the non-cationic lipid is a negatively charged (anionic) lipid. [00506] Exemplary non-cationic lipids include, but are not limited to, distearoyl-sn-glycero- phosphoethanolamine, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane- 1 -carboxylate (DOPE- mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), monomethyl-phosphatidylethanolamine (such as 16-O-monomethyl PE), dimethyl- phosphatidylethanolamine (such as 16-O-dimethyl PE), 18-1- trans PE, l-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), hydrogenated soy phosphatidylcholine (HSPC), egg phosphatidylcholine (EPC), dioleoylphosphatidylserine (DOPS), sphingomyelin (SM), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphatidylglycerol (DMPG), distearoylphosphatidylglycerol (DSPG), dierucoylphosphatidylcholine (DEPC), palmitoyloleyolphosphatidylglycerol (POPG), dielaidoylphosphatidylethanolamine (DEPE), 1,2-dilauroyl- sn-glycero-3-phosphocholine (DLPC), Sodium 1,2- ditetradecanoyl-sn-glycero-3-phosphate (DMPA), phosphatidylcholine (lecithin), phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), phosphatidylethanolamine (cephalin), cardiolipin, phosphatidic acid, cerebrosides, dicetylphosphate, lysophosphatidylcholine, dilinoleoylphosphatidylcholine, or mixtures thereof. It is understood that other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids can also be used. The acyl groups in these lipids are preferably acyl groups derived from fatty acids having C10-C24 carbon chains, e.g., lauroyl, myristoyl, paimitoyl, stearoyl, or oleoyl. Additional exemplary lipids, in certain embodiments, include, without limitation, those described in Kim et al. (2020) dx.doi.org/10.1021/acs.nanolett.0c01386, the entire contents of which a e incorporated by reference herein for all purposes. Such lipids include, in some embodiments, plant lipids found to improve liver transfection with mRNA (e.g., DGTS).
[00507] In some embodiments, the lipid-based carrier (or lipid nanoformulation) may comprise a combination of distearoylphosphatidylcholine/cholesterol, dipalmitoylphosphatidy Icholine/cholesterol, dimyry stoylphosphatidylcholine/cholesterol, 1 ,2- Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/cholesterol, or egg sphingomyelin/cholesterol.
[00508] Other examples of suitable non-cationic lipids include, without limitation, nonphosphorous lipids such as, e.g., stearylamine, dodecylamine, hexadecylamine, acetyl palmitate, glycerol ricinoleate, hexadecyl stearate, isopropyl myristate, amphoteric acrylic polymers, triethanolamine-lauryl sulfate, alkyl-aryl sulfate polyethyloxylated fatty acid amides, dioctadecyl dimethyl ammonium bromide, ceramide, sphingomyelin, and the like. Other noncationic lipids are described in WO 2017/099823 or US 2018/0028664, the entire contents of each of which are incorporated by reference herein for all purposes.
[00509] In one embodiment, the lipid-based carrier (or lipid nanoformulation) further comprises one or more non-cationic lipid that is oleic acid or a compound of Formula I, II, or IV of US 2018/0028664, the entire contents of which are incorporated by reference herein for all purposes. [00510] The non-cationic lipid content can be, for example, 0-30% (mol) of the total lipid components present. In some embodiments, the non-cationic lipid content is 5-20% (mol) or 10- 15% (mol) of the total lipid components present.
[00511] In some embodiments, the lipid-based carrier (or lipid nanoformulation) further comprises a neutral lipid, and the molar ratio of an ionizable lipid to a neutral lipid ranges from about 2:1 to about 8:1 (e.g., about 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, or 8:1).
[00512] In some embodiments, the lipid-based carrier (or lipid nanoformulation) does not include any phospholipids.
[00513] In some embodiments, the lipid-based carrier (or lipid nanoformulation) can further include one or more phospholipids, and optionally one or more additional molecules of similar molecular shape and dimensions having both a hydrophobic moiety and a hydrophilic moiety (e.g., cholesterol).
4.9.1.3 Structural Lipids
[00514] The lipid-based carrier (or lipid nanoformulation) described herein may further comprise one or more structural lipids. As used herein, the term “structural lipid” refers to sterols (e.g., cholesterol) and also to lipids containing sterol moieties. [00515] Incorporation of structural lipids in the lipid nanoparticle may help mitigate aggregation of other lipid in the particle. Structural lipids can be selected from the group including but not limited to, cholesterol or cholesterol derivative, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, tomatine, ursolic acid, alpha-tocopherol, hopanoids, phytosterols, steroids, and mixtures thereof. In some embodiments, the structural lipid is a sterol. In certain embodiments, the structural lipid is a steroid. In certain embodiments, the structural lipid is cholesterol. In certain embodiments, the structural lipid is an analog of cholesterol. In certain embodiments, the structural lipid is alpha-tocopherol.
[00516] In some embodiments, structural lipids may be incorporated into the lipid-based carrier at molar ratios ranging from about 0.1 to 1.0 (cholesterol phospholipid).
[00517] In some embodiments, sterols, when present, can include one or more of cholesterol or cholesterol derivatives, such as those described in WO 2009/127060 or US 2010/0130588, the entire contents of each of which are incorporated by reference herein for all purposes. Additional exemplary sterols include phytosterols, including those described in Eygeris et al. (2020), Nano Lett. 2020;20(6):4543-4549, the entire contents of which are incorporated by reference herein for all purposes.
[00518] In some embodiments, the structural lipid is a cholesterol derivative. Non-limiting examples of cholesterol derivatives include polar analogues such as 5a-cholestanol, 53- coprostanol, cholesteryl-(2’-hydroxy)-ethyl ether, cholesteryl-(4'- hydroxy )-butyl ether, and 6- ketocholestanol; non-polar analogues such as 5a-cholestane, cholestenone, 5a-cholestanone, 5p- cholestanone, and cholesteryl decanoate; and mixtures thereof. In some embodiments, the cholesterol derivative is a polar analogue, e.g., cholesteryl-(4'-hydroxy)-butyl ether. Exemplary cholesterol derivatives are described in WO 2009/127060 and US 2010/0130588, the entire contents of each of which are incorporated by reference herein for all purposes.
[00519] In some embodiments, the lipid-based carrier (or lipid nanoformulation) further comprises sterol in an amount of 0-50 mol% (e.g., 0-10 mol %, 10-20 mol %, 20-50 mol%, 20-30 mol %, 30-40 mol %, or 40-50 mol %) of the total lipid components.
4.9.1.4 Polymers and Polyethylene Glycol (PEG) - Lipids
[00520] In some embodiments, the lipid-based carrier (or lipid nanoformulation) may include one or more polymers or co-polymers, e.g., poly(lactic-co-glycolic acid) (PFAG) nanoparticles.
[00521] In some embodiments, the lipid-based carrier (or lipid nanoformulation) may include one or more polyethylene glycol (PEG) lipid. Examples of useful PEG-lipids include, but are not limited to, l,2-Diacyl-sn-Glyccro-3- Phosphocthanolaminc-N- [Methoxy (Polyethylene glycol)- 350] (mPEG 350 PE); 1,2-Diacyl-sn- Glycero-3-Phosphoethanolamine-N-[Methoxy(Polyethylene glycol)-550] (mPEG 550 PE); 1,2- Diacyl-sn-Glycero-3-Phosphoethanolamine-N- [Methoxy(Polyethylene glycol)-750] (mPEG 750 PE); l,2-Diacyl-sn-Glycero-3- Phosphoethanolamine-N-[Methoxy(Polyethylene glycol)-1000] (mPEG 1000 PE); 1,2-Diacyl-sn- Glycero-3-Phosphoethanolamine-N-[Methoxy(Polyethylene glycol)-2000] (mPEG 2000 PE); 1,2- Diacyl-sn-Glycero-3-Phosphoethanolamine-N- [Methoxy(Polyethylene glycol)-3000] (mPEG 3000 PE); l,2-Diacyl-sn-Glycero-3- Phosphoethanolamine-N- [Methoxy (Polyethylene glycol)- 5000] (mPEG 5000 PE); N-Acyl- Sphingosine- l-[Succinyl(Methoxy Polyethylene Glycol) 750] (mPEG 750 Ceramide); N-Acyl- Sphingosine- l-[Succinyl(Methoxy Polyethylene Glycol) 2000] (mPEG 2000 Ceramide); and N- Acyl-Sphingosine-l-[Succinyl(Methoxy Polyethylene Glycol) 5000] (mPEG 5000 Ceramide). In some embodiments, the PEG lipid is a polyethyleneglycoldiacylglycerol (i.e., polyethyleneglycol diacylglycerol (PEG-DAG), PEG-cholesterol, or PEG- DMB) conjugate.
[00522] In some embodiments, the lipid-based carrier (or nanoformulation) includes one or more conjugated lipids (such as PEG-conjugated lipids or lipids conjugated to polymers described in Table 5 of WO 2019/217941, the entire contents of which are incorporated by reference herein for all purposes). In some embodiments, the one or more conjugated lipids is formulated with one or more ionic lipids (e.g., non-cationic lipid such as a neutral or anionic, or zwitterionic lipid); and one or more sterols (e.g., cholesterol).
[00523] The PEG conjugate can comprise a PEG-dilaurylglycerol (C12), a PEG- dimyristylglycerol (C14), a PEG-dipalmitoylglycerol (C16), a PEG-disterylglycerol (C18), PEG- dilaurylglycamide (C12), PEG-dimyristylglycamide (C14), PEG-dipalmitoylglycamide (C16), and PEG-disterylglycamide (Cl 8).
[00524] In some embodiments, conjugated lipids, when present, can include one or more of PEG-diacylglycerol (DAG) (such as l-(monomethoxy-polyethyleneglycol)-2,3- dimyristoylglycerol (PEG-DMG)), PEG-dialkyloxypropyl (DAA), PEG-phospholipid, PEG- ceramide (Cer), a pegylated phosphatidylethanoloamine (PEG-PE), PEG succinate diacylglycerol (PEGS-DAG) (such as 4-0-(2',3'-di(tetradecanoyloxy)propyl-l-0-(w- methoxy(polyethoxy)ethyl) butanedioate (PEG-S-DMG)), PEG dialkoxypropylcarbam, N- (carbonyl-methoxypolyethylene glycol 2000)- 1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine sodium salt, and those described in Tabic 2 of WO 2019/051289 (the entire contents of which arc incorporated by reference herein for all purposes), and combinations of the foregoing.
[00525] Additional exemplary PEG-lipid conjugates are described, for example, in US 5,885,613, US 6,287,591, US 2003/0077829, US 2003/0077829, US 2005/0175682, US 2008/0020058, US 2011/0117125, US 2010/0130588, US 2016/0376224, US 2017/0119904, US 2018/0028664, and WO 2017/099823, the entire contents of each of which are incorporated by reference herein for all puiposes.
[00526] In some embodiments, the PEG-lipid is a compound of Formula III, III-a-I, III-a-2, III- b-1, III-b-2, or V of US 2018/0028664, which is incorporated herein by reference in its entirety. In some embodiments, the PEG-lipid is of Formula II of US 2015/0376115 or US 2016/0376224, the entire contents of each of which are incorporated by reference herein for all purposes. In some embodiments, the PEG-DAA conjugate can be, for example, PEG-dilauryloxypropyl, PEG- dimyristyloxypropyl, PEG-dipalmityloxypropyl, or PEG-distearyloxypropyl. In some embodiments, the PEG-lipid includes one of the following:
[00527] In some embodiments, lipids conjugated with a molecule other than a PEG can also be used in place of PEG-lipid. For example, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (such as ATTA-lipid conjugates), and cationic-polymer lipid (GPL) conjugates can be used in place of or in addition to the PEG-lipid.
[00528] Exemplary conjugated lipids, e.g., PEG-lipids, (POZ)-lipid conjugates, ATTA-lipid conjugates and cationic polymer-lipids, include those described in Table 2 of WO 2019/051289A9, the entire contents of which arc incorporated by reference herein for all purposes.
[00529] In some embodiments, the conjugated lipid (e.g., the PEGylated lipid) can be present in an amount of 0-20 mol% of the total lipid components present in the lipid-based carrier (or lipid nanoformulation). In some embodiments, the conjugated lipid (e.g., the PEGylated lipid) content is 0.5-10 mol% or 2-5 mol% of the total lipid components.
[00530] When needed, the lipid-based carrier (or lipid nanoformulation) described herein may be coated with a polymer layer to enhance stability in vivo (e.g., sterically stabilized LNPs).
[00531] Examples of suitable polymers include, but are not limited to, poly(ethylene glycol), which may form a hydrophilic surface layer that improves the circulation half-life of liposomes and enhances the amount of lipid nanoformulations (e.g., liposomes or LNPs) that reach therapeutic targets. See, e.g., Working et al. J Pharmacol Exp Ther, 289: 1128-1133 (1999); Gabizon et al., J Controlled Release 53: 275-279 (1998); Adlakha Hutcheon et al., Nat Biotechnol 17: 775-779 (1999); and Koning et al., Biochim Biophys Acta 1420: 153-167 (1999), the entire contents of each of which are incorporated by reference herein for all purposes.
4.9.1.5 Percentages of Lipid Nanoformulation Components
[00532] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises one of more of the RNA agents described herein (e.g., RNAi agents, dsRNA agents, sense strands, antisense strands (or a conjugate comprising the same)) (or a vector comprising any of the foregoing), optionally a non-cationic lipid (e.g., a phospholipid), a sterol, a neutral lipid, and optionally conjugated lipid (e.g., a PEGylated lipid) that inhibits aggregation of particles. In some embodiments, the lipid-based carrier (or lipid nanoformulation) further comprises an RNA agent (e.g., an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing))). The amounts of these components can be varied independently and to achieve desired properties. For example, in some embodiments, the ionizable lipid including the lipid compounds described herein is present in an amount from about 20 mol% to about 100 mol% (e.g., 20-90 mol%, 20-80 mol%, 20-70 mol%, 25-100 mol%, 30-70 mol%, 30-60 mol%, 30-40 mol%, 40-50 mol%, or 50-90 mol%) of the total lipid components; a non-cationic lipid (e.g., phospholipid) is present in an amount from about 0 mol% to about 50 mol% (e.g., 0-40 mol%, 0- 30 mol%, 5-50 mol%, 5-40 mol%, 5-30 mol%, or 5-10 mol%) of the total lipid components, a conjugated lipid (e.g., a PEGylated lipid) in an amount from about 0.5 mol% to about 20 mol% (e.g., 1-10 mol% or 5-10%) of the total lipid components, and a sterol in an amount from about 0 mol % to about 60 mol% (e.g., 0-50 mol%, 10-60 mol%, 10-50 mol%, 15-60 mol%, 15-50 mol%, 20-50 mol%, 20-40 mol%) of the total lipid components, provided that the total mol% of the lipid component does not exceed 100%.
[00533] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises about 25-100 mol% of the ionizable lipid including the lipid compounds described herein, about 0-50 mol% phospholipid, about 0-50 mol% sterol, and about 0-10 mol% PEGylated lipid.
[00534] In some embodiments, the lipid-based earner comprises an RNA agent described herein e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) that is formulated in a lipid nanoparticle, wherein the lipid nanoparticle comprises about 25-100 mol% of the ionizable lipid including the lipid compounds described herein, about 0-50 mol% phospholipid, about 0-50 mol% sterol, and about 0-10 mol% PEGylated lipid. In some embodiments, the encapsulation efficiency of the RNA agent may be at least 70%.
[00535] In one embodiment, the lipid-based carrier (or lipid nanoformulation) comprises about 25-100 mol% of the ionizable lipid including the lipid compounds described herein; about 0-40 mol% phospholipid (e.g., DSPC), about 0-50 mol% sterol (e.g., cholesterol), and about 0-10 mol% PEGylated lipid.
[00536] In some embodiments, the lipid-based carrier comprises an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) that is formulated in a lipid nanoparticle, wherein the lipid nanoparticle comprises about 25-100 mol% of the ionizable lipid including the lipid compounds described herein; about 0-40 mol% phospholipid (e.g., DSPC), about 0-50 mol% sterol (e.g., cholesterol), and about 0-10 mol% PEGylated lipid. In some embodiments, the encapsulation efficiency of the RNA agent may be at least 70%.
[00537] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises about 30-60 mol% (e.g., about 35-55 mol%, or about 40-50 mol%) of the ionizable lipid including the lipid compounds described herein, about 0-30 mol% (e.g., 5-25 mol%, or 10-20 mol%) phospholipid, about 15-50 mol% (e.g., 18.5-48.5 mol%, or 30-40 mol%) sterol, and about 0-10 mol% (e.g., 1-5 mol%, or 1.5-2.5 mol%) PEGylated lipid. [00538] In some embodiments, the lipid-based carrier comprises an RNA agent described herein (e.g., RNAi agent, dsRNA agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) that is formulated in a lipid nanoparticle, wherein the lipid nanoparticle comprises about 30-60 mol% (e.g., about 35-55 mol%, or about 40- 50 mol%) of the ionizable lipid including the lipid compounds described herein, about 0-30 mol% (e.g., 5-25 mol%, or 10-20 mol%) phospholipid, about 15-50 mol% (e.g., 18.5-48.5 mol%, or 30- 40 mol%) sterol, and about 0-10 mol% (e.g., 1-5 mol%, or 1.5-2.5 mol%) PEGylated lipid. In some embodiments, the encapsulation efficiency of the RNA agent may be at least 70%.
[00539] In some embodiments, molar ratios of ionizable lipid/sterol/phospholipid (or another structural lipid)/PEG-lipid/additional components is varied in the following ranges: ionizable lipid (25-100%); phospholipid (DSPC) (0-40%); sterol (0-50%); and PEG lipid (0-5%).
[00540] In some embodiments, the lipid-based carrier comprises an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) that is formulated in a lipid nanoparticle, wherein the lipid nanoparticle comprises molar ratios of ionizable lipid/sterol/phospholipid (or another structural lipid)/PEG-lipid/additional components in the following ranges: ionizable lipid (25-100%); phospholipid (DSPC) (0-40%); sterol (0-50%); and PEG lipid (0-5%). In some embodiments, the encapsulation efficiency of the RNA agent may be at least 70%.
[00541] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises, by mol% or wt% of the total lipid components, 50-75% ionizable lipid (including the lipid compound as described herein), 20-40% sterol (e.g., cholesterol or derivative), 0 to 10% non-cationic-lipid, and 1-10% conjugated lipid (e.g., the PEGylated lipid).
[00542] In some embodiments, the lipid-based carrier comprises an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) that is formulated in a lipid nanoparticle, wherein the lipid nanoparticle comprises, by mol% or wt% of the total lipid components, 50-75% ionizable lipid (including the lipid compound as described herein), 20-40% sterol (e.g., cholesterol or derivative), 0 to 10% non-cationic-lipid, and 1-10% conjugated lipid (e.g., the PEGylated lipid). In some embodiments, the encapsulation efficiency of the RNA agent may be at least 70%. [00543] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises (i) an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing); (ii) a cationic lipid comprising from 50 mol% to 65 mol% of the total lipid present in the lipid-based carrier; (iii) a non-cationic lipid comprising a mixture of a phospholipid and a cholesterol derivative thereof, wherein the phospholipid comprises from 3 mol% to 15 mol% of the total lipid present in the lipid-based carrier and the cholesterol or derivative thereof comprises from 30 mol% to 40 mol% of the total lipid present in the lipid-based carrier; and (iv) a conjugated lipid comprising 0.5 mol% to 2 mol% of the total lipid present in the particle.
[00544] In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises (i) an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing); (ii) a cationic lipid comprising from 50 mol % to 85 mol % of the total lipid present in the lipid-based carrier; (iii) a non-cationic lipid comprising from 13 mol % to 49.5 mol % of the total lipid present in the lipid-based earner; and (d) a conjugated lipid comprising from 0.5 mol % to 2 mol % of the total lipid present in the lipid-based earner.
[00545] In some embodiments, the phospholipid component in the mixture may be present from 2 mol% to 20 mol%, from 2 mol% to 15 mol%, from 2 mol% to 12 mol%, from 4 mol% to 15 mol%, from 4 mol% to 10 mol%, from 5 mol% to 10 mol%, (or any fraction of these ranges) of the total lipid components. In some embodiments, the lipid-based carrier (or lipid nanoformulation) is phospholipid-free.
[00546] In some embodiments, the sterol component (e.g. cholesterol or derivative) in the mixture may comprise from 25 mol% to 45 mol%, from 25 mol% to 40 mol%, from 25 mol% to 35 mol%, from 25 mol% to 30 mol%, from 30 mol% to 45 mol%, from 30 mol% to 40 mol%, from 30 mol% to 35 mol%, from 35 mol% to 40 mol%, from 27 mol% to 37 mol%, or from 27 mol% to 35 mol% (or any fraction of these ranges) of the total lipid components.
[00547] In some embodiments, the non-ionizable lipid components in the lipid-based carrier (or lipid nanoformulation) may be present from 5 mol% to 90 mol%, from 10 mol% to 85 mol%, or from 20 mol% to 80 mol% (or any fraction of these ranges) of the total lipid components.
[00548] The ratio of total lipid components to the RNA agent (e.g., an encapsulated agent such as an RNA agent described herein (e.g., RNAi agent, double stranded RNA (dsRNA) agent, sense strand, antisense strand (or a conjugate comprising the same)) (or a vector comprising any of the foregoing) can be varied as desired. For example, the total lipid components to the RNA agent (mass or weight) ratio can be from about 10:1 to about 30:1. In some embodiments, the total lipid components to the RNA agent ratio (mass/mass ratio; w/w ratio) can be in the range of from about 1:1 to about 25:1, from about 10:1 to about 14:1, from about 3:1 to about 15:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or about 6:1 to about 9:1. The amounts of total lipid components and the RNA agent can be adjusted to provide a desired N/P ratio, for example, N/P ratio of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or higher. Generally, the lipid-based carrier (or lipid nanoformulation’s) overall lipid content can range from about 5 mg/ml to about 30 mg/mL. Nitrogemphosphate ratios (N:P ratio) is evaluated at values between 0.1 and 100.
[00549] The efficiency of encapsulation of an RNA agent (e.g., an RNA agent described herein), describes the amount of agent that is encapsulated or otherwise associated with a lipid nanoformulation (e.g., liposome or LNP) after preparation, relative to the initial amount provided. The encapsulation efficiency is desirably high (e.g., at least 70%. 80%. 90%. 95%, close to 100%). The encapsulation efficiency may be measured, for example, by comparing the amount of agent in a solution containing the liposome or LNP before and after breaking up the liposome or LNP with one or more organic solvents or detergents. An anion exchange resin may be used to measure the amount of free agent in a solution. Fluorescence may be used to measure the amount of free agent in a solution. For the lipid-based carrier (or lipid nanoformulation) described herein, the encapsulation efficiency of a protein and/or nucleic acid may be at least 50%, for example 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the encapsulation efficiency may be at least 70%. In some embodiments, the encapsulation efficiency may be at least 80%. In some embodiments, the encapsulation efficiency may be at least 90%. In some embodiments, the encapsulation efficiency may be at least 95%.
4.10 Pharmaceutical Compositions
[00550] In one aspect, provided herein are pharmaceutical compositions comprising any one more agent (e.g., RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand) (see, e.g., §§ 4.2, 4.3)); a vector described herein (see, e.g., § 4.7); a conjugate described herein (see, e.g., § 4.4); a carrier described herein (see, e.g., § 4.9); and/or a cell described herein (see, e.g., § 4.8); or any combination thereof; and a pharmaceutically acceptable excipient (see, e.g., Remington’s Pharmaceutical Sciences (1990) Mack Publishing Co., Easton, PA, the entire contents of which is incorporated by reference herein for all purposes). [00551] In one aspect, also provided herein are methods of making pharmaceutical compositions described herein comprising providing any one or more agent (e.g., RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand) (see, e.g., §§ 4.2, 4.3)); a vector described herein (see, e.g., § 4.7); a conjugate described herein (see, e.g., § 4.4); a carrier described herein (see, e.g., § 4.9); and/or a cell described herein (see, e.g., § 4.8); and formulating it into a pharmaceutically acceptable composition by the addition of one or more pharmaceutically acceptable excipient.
[00552] Also provided herein are pharmaceutical compositions comprising any one or more agent (e.g., RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand)); vector described herein; conjugate described herein; carrier described herein; and/or cell described herein, wherein the pharmaceutical composition lacks a predetermined threshold amount or a detectable amount of a process impurity or contaminant, e.g., lacks a predetermined threshold amount or a detectable amount of a process-related impurity such as host cell proteins, host cell DNA, or a cell culture component (e.g., inducers, antibiotics, or media components); a product-related impurity (e.g., precursors, fragments, aggregates, degradation products); or a contaminant, e.g., endotoxin, bacteria, viral contaminant.
[00553] Acceptable excipients (e.g., carriers and stabilizers) are preferably nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, or other organic acids; antioxidants including ascorbic acid or methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol;or m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, or other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).
[00554] A pharmaceutical composition may be formulated for any route of administration to a subject. Non-limiting embodiments include parenteral administration, such as intramuscular, intradermal, subcutaneous, transcutaneous, or mucosal.
[00555] In one embodiment, the pharmaceutical composition is formulated for administration by intramuscular, intradermal, or subcutaneous injection. In one embodiment, the pharmaceutical composition is formulated for administration by intramuscular injection. In one embodiment, the pharmaceutical composition is formulated for administration by intradermal injection. In one embodiment, the pharmaceutical composition is formulated for administration by subcutaneous injection. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions. The injectables can contain one or more excipients. Exemplary excipients include, for example, water, saline, dextrose, glycerol or ethanol. In addition, if desired, the pharmaceutical compositions to be administered can also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, or other such agents, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate or cyclodextrins. In some embodiments, the pharmaceutical composition is formulated in a single dose. In some embodiments, the pharmaceutical compositions if formulated as a multi-dose. [00556] Pharmaceutically acceptable excipients used in the parenteral preparations described herein include for example, aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents or other pharmaceutically acceptable substances. Examples of aqueous vehicles, which can be incorporated in one or more of the formulations described herein, include sodium chloride injection, Ringer’s injection, isotonic dextrose injection, sterile water injection, dextrose or lactated Ringer’s injection. Nonaqueous parenteral vehicles, which can be incorporated in one or more of the formulations described herein, include fixed oils of vegetable origin, cottonseed oil, corn oil, sesame oil or peanut oil. Antimicrobial agents in bacteriostatic or fungistatic concentrations can be added to the parenteral preparations described herein and packaged in multiple-dose containers, which include phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride or benzethonium chloride. Isotonic agents, which can be incorporated in one or more of the formulations described herein, include sodium chloride or dextrose. Buffers, which can be incorporated in one or more of the formulations described herein, include phosphate or citrate. Antioxidants, which can be incorporated in one or more of the formulations described herein, include sodium bisulfate. Local anesthetics, which can be incorporated in one or more of the formulations described herein, include procaine hydrochloride. Suspending and dispersing agents, which can be incorporated in one or more of the formulations described herein, include sodium carboxymethylcelluose, hydroxypropyl methylcellulose or polyvinylpyrrolidone. Emulsifying agents, which can be incorporated in one or more of the formulations described herein, include Polysorbate 80 (TWEEN® 80). A sequestering or chelating agent of metal ions, which can be incorporated in one or more of the formulations described herein, is EDTA. Pharmaceutical carriers, which can be incorporated in one or more of the formulations described herein, also include ethyl alcohol, polyethylene glycol or propylene glycol for water miscible vehicles; orsodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.
[00557] The precise dose to be employed in a pharmaceutical composition will also depend on the route of administration, and the seriousness of the condition caused by it, and should be decided according to the judgment of the practitioner and each subject’s circumstances. For example, effective doses may also vary depending upon means of administration, target site, physiological state of the subject (including age, body weight, and health), other medications administered, or whether therapy is prophylactic or therapeutic. Therapeutic dosages are preferably titrated to optimize safety and efficacy.
4.11 Exemplary Somatic Mutations and Detection Methods
[00558] Several aspects and embodiments described herein comprise a step of determining (or having determined) the presence or absence of one or more somatic mutation (e.g., in DNA, RNA, or protein from a sample) (see, e.g., §§ 4.15.1, 4.15.2, 4.15.3, 4.15.4). Methods of detecting somatic mutations are known in the ail. See, e.g., Dou, Yanmei et al. “Detecting Somatic Mutations in Normal Cells.” Trends in genetics: TIG vol. 34,7 (2018): 545-557. doi: 10.1016/j .tig.2018.04.003; Huang August Yue, Lee Eunjung Alice, “Identification of Somatic Mutations From Bulk and Single-Cell Sequencing Data,” Frontiers in Aging, Vol 2 (2022) DOI=10.3389/fragi.2021.800380; Muyas, F., Sauer, C.M., Valle-Inclan, J.E. et al. De novo detection of somatic mutations in high-throughput single-cell profiling data sets. Nat Biotechnol (2023). https://doi.org/l0.1038/s4l587-023-0l 863-z, Martincorena, T. et al., Cell 2017, 171, 1029- 1041. c21, and the NBR algorithm described by Rhcinbay, E. ct al., Nature, 2020, 578, 102-111; the entire contents of each of which are incorporated herein by reference for all purposes. [00559] In some embodiments, the somatic mutation is a gain of function mutation. In some embodiments, the somatic mutation is a loss of function mutation. In some embodiments, the somatic mutation comprises one or more nucleotide substitution, addition, or deletion. In some embodiments, the somatic mutation is a single nucleotide substitution. In some embodiments, the somatic mutation is non-synonymous. In some embodiments, the somatic mutation is synonymous. In some embodiments, the somatic mutation is a single nucleotide polymorphism (SNP). In some embodiments, the somatic mutation is a synonymous SNP. In some embodiments, the somatic mutation is a non-synonymous SNP. In some embodiments, the somatic mutation changes the amino acid sequence of the encoded protein (e.g., the amino acid sequence of the encoded protein comprises one or more amino acid substitution, deletion, addition, etc.). In some embodiments, the somatic mutation is a truncating mutation. In some embodiments, the somatic mutation is an INDEL mutation. In some embodiments, the somatic mutation is an insertion. In some embodiments, the somatic mutation is a deletion. In some embodiments, the somatic mutation comprises a frameshift mutation. In some embodiments, the somatic mutations comprises a missense mutation. In some preferred embodiments, the somatic mutations comprises a missense mutation. In some embodiments, the somatic mutation does not result in a change in the amino acid sequence of the encoded protein. In some embodiments, the somatic mutation comprises a nonsense mutation. In some embodiments, the somatic mutation can result in the generation of a stop codon (e.g., resulting in a truncated variant of the encoded protein). In some embodiments, the somatic mutation effects splicing. In some embodiments, the somatic mutation is in a splice site. In specific embodiments, the somatic mutation disrupts expression of the protein encoded by the gene. In specific embodiments, the somatic mutation truncates the protein encoded by the gene. [00560] Table 2 provides exemplary specific somatic mutations of each of genes BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6, that may be utilized (e.g., analyzed, detected, etc.) in any one of the method described herein. The somatic mutations are denoted according to UCSC Genome Browser assembly ID: hg38 (Sequencing/Assembly provider ID: Genome Reference Consortium Human GRCh38.pl4 (GCA_000001405.29); Assembly date: Dec. 2013 initial release; June 2022 patch release 14; Assembly accession: GCA_000001405.29; NCBI Genome ID: 51 (Homo sapiens (human)); NCBI Assembly ID: GCF_000001405.40 (GRCh38.pl4, GCA_000001405.29) BioProjcct ID: PRJNA31257). In specific embodiments, the one or more somatic mutation that is utilized (e.g., analyzed, detected, etc.) in any method described herein is one set forth in Table 2.
Table 2. Exemplary Somatic Mutations in BTG1, CDKN1B, DUSP2, IGLL5, and KLHL6.
2
[00561] The methods can be carried out utilizing more than one sample from the subject. In some embodiments, each of the samples is from a different biopsy. In some embodiments, each of the samples is from the same biopsy (e.g., a different microdissection of the same biopsy). In some embodiments, each of the samples is a different blood sample. In some embodiments, at least one of the samples is a solid tissue sample and one of the samples is a fluid sample (e.g., a blood sample, a urine sample, etc.). In some embodiments, the sample comprises a population of immune cells. In some embodiments, the sample comprises B cells.
[00562] In some embodiments, the methods further comprises quantifying the level of one or more somatic mutations in the sample. Techniques suitable for quantifying the level of one or more somatic mutations are known in the art, and include, for example, the use of nanopore sequencing, qPCR, RT-qPCR, DNA/RNA microarrays and analysis of DNA sequencing data, particularly next-generation sequencing (e.g., Illumina dye sequencing and Ion Torrent semiconductor sequencing) data. The quantification of the level of the somatic mutation can include an assessment of any one or more of e.g., the copy number of one or more somatic mutations in a given sample, the concentration of DNA molecules comprising the somatic mutation in a given sample, the concentration of mRNA molecules comprising the somatic mutation in a given sample, or the concentration of a protein comprising the one or more amino acid variation encoded by the somatic mutation in a given sample.
[00563] In some embodiments, the level of the somatic mutation e.g., as assessed by the copy number of a mutation, the concentration of DNA with the mutation, the concentration of mRNA with the mutation and/or the concentration of protein with the mutation) is compared to the level of the somatic mutation in a reference sample obtained from a healthy subject (e.g., a sample obtained from a subject that does not have a reference disease), a reference sample obtained from a subject known to have a reference disease, a reference sample obtained from a subject that has not been diagnosed with a reference disease, and/or a reference sample obtained from the same subject (e.g., a different type of sample).
[00564] In some embodiments the reference sample is from a healthy subject. In some embodiments, if the level of somatic mutation in the given sample from the subject is higher than the level in the reference sample, this indicates that the subject has a reference disease, has an increased risk of developing a reference disease, has an increased risk of developing a more severe form of a reference disease, and/or has an increased risk of developing a disease or condition associated with the reference disease. In some embodiments, if the level of the somatic mutation in a given biological sample from the subject is comparable to, or lower than, the level in the reference sample obtained from a healthy subject this indicates that the subject does not have the reference disease, does not have an increased risk of developing the reference disease, does not have an increased risk of developing a more severe form of the reference disease and/or does not have an increased risk of developing a disease or condition associated with the reference disease. [00565] In some embodiments the reference sample is from a subject known to have a reference disease. In some embodiments, if the level of the somatic mutation in the given sample from the subject is comparable to, or higher than, the level in the reference sample, this indicates that the subject has a reference disease, has an increased risk of developing a reference disease, has an increased risk of developing a more severe form of a reference disease and/or an increased risk of developing a disease associated with the reference disease. In some embodiments, the level of the somatic mutation in the given sample from the subject is lower than the level in the reference sample this may indicate that the subject docs not have the reference disease, docs not have an increased risk of developing the reference disease, does not have an increased risk of developing a more severe form of the reference disease and/or does not have an increased risk of developing a disease associated with the reference disease.
[00566] In some embodiments the reference sample is from the same subject as the given sample. In some embodiments, the reference sample from the same subject is obtained from a different region of the body of the subject, and/or obtained from the subject at a different time point to the given sample. For example, the reference sample may be obtained from the subject, 1 to 6 months earlier, 6 to 12 months earlier, 1 to 2 year’s earlier or 2 to 3 years earlier than the given sample obtained from the subject.
[00567] In some embodiments wherein the reference sample is obtained from the same subject but at an earlier time point to the given sample. In some embodiments, if the level of one or more somatic mutation in the given sample is higher than the level in the reference sample, this may indicate that the subject has an increased risk of developing a reference disease, has an increased risk of developing a more severe form of a reference disease and/or has an increased risk of developing a disease associated with the reference disease compared to the earlier time point when the reference sample was obtained from the subject. Alternatively, in some embodiments, if the level of the somatic mutation in the given sample is higher than the level in the reference sample, this may indicate that the subject has developed a reference disease, has developed a more severe form of a reference disease and/or has developed a disease associated with the reference disease.
[00568] In some embodiments, if the level of each one or more somatic mutations in the given sample from the subject is comparable to the level in the reference sample this may indicate that the subject has not developed a subject disease, has not developed a more severe form of a subject disease, and/or has not developed a disease associated with the subject disease. In some embodiments, if the level of each one or more somatic mutations in the given sample from the subject is lower than the level in the reference sample this may indicate that the subject has a decreased risk of developing a subject disease, has a decreased risk of developing a more severe form of a subject disease and/or has a decreased risk of developing a disease associated with the subject disease compared to the earlier time point when the reference sample was obtained from the subject. In some embodiments, if the level of each one or more somatic mutations in the given sample from the subject is lower than the level in the reference sample this may indicate that the subject has a subject that is in remission, has a more severe form of a subject disease that is in remission and/or has a disease associated with a subject that is in remission.
[00569] Several methods described herein comprise a cell or a population of cells (e.g., a cell or population of cells in vitro, in vivo, in a subject, etc.) In some embodiments, the cell is an immune cell. Exemplary immune cells include, T cells (e.g., CD4+ T cells, CD8+ T cells), B-cells, tumor infiltrating lymphocytes (e.g., tumor infiltrating T cells (e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells)). In some embodiments, the cell is a T-cell. In some embodiments, the cell is a tumor infiltrating lymphocyte. In some embodiments, the cell is a tumor infiltrating T cell. In some embodiments, the cell is a tumor infiltrating CD4+ T cell. In some embodiments, the cell is a CD8+ tumor infiltrating T cell. In some embodiments, the cell is B-cell.
4.12 Exemplary Samples and Methods of Processing
[00570] Several methods described herein comprise the use or obtaining of a sample from a subject (see, e.g., §§ 4.15.1, 4.15.2, 4.15.3, 4.15.4). In some embodiments, the sample is a blood, red blood cell, or other liquid samples of biological origin (e.g., whole-blood, red blood cells (e.g., isolated from whole blood), peripheral blood mononuclear cells (PBMCs), serum, plasma, urine, saliva, amniotic fluid, stool, synovial fluid, etc.), a nasopharyngeal swab, a solid tissue sample such as a biopsy (or cells derived therefrom and the progeny thereof), tissue culture (or cells derived therefrom and the progeny thereof), or cell culture (or cells derived therefrom and the progeny thereof). In one embodiment, the sample is a biopsy. In some embodiments, the sample is a bowel biopsy. In one embodiment, the sample is a blood (e.g., whole blood sample). Methods of obtaining various types of samples are known in the art and are within the routine abilities of a clinical practitioner.
[00571] In one embodiment, the sample comprises cells. In embodiments, wherein the sample comprises cells, the sample can comprise e.g., at least 10, at least 100, at least 1000, at least 10,000, at least 104, at least 105, or at least 106 cells. For example, about 10 to about 106 cells, about 100 to about 105, about 500 to about 104, about 10 to about 50,000, or about 1000 to about 50,000 (for example, about 1000 to about 10,000, about 1000 to about 20,000, about 1000 to about 30,000, or about 1000 to about 40,000 cells). In exemplary embodiments, the sample comprises DNA, RNA and/or protein derived from about 10 to about 50,000 cells (for example, about 10 to about 50,000 cells) of the subject. In some embodiments, the sample contains about a single cell. In some embodiments, a single cell is isolated from the sample for utilization in a method described herein. In exemplary embodiments, the sample comprises DNA, RNA and/or protein derived from about a single cell.
[00572] In certain embodiments, the method comprises detecting and/or quantifying one or more somatic mutations derived from different cell clones contained in a given sample obtained from a subject. In certain embodiments, the method comprises detecting and/or quantifying one or more somatic mutations derived from different cell clones contained in different samples each obtained from a subject.
[00573] In some embodiments, the sample comprises DNA (e.g., genomic DNA), RNA (e.g., mRNA), or protein. For example, the sample may comprise DNA or RNA extracted from cells present in a blood sample or biopsy sample obtained from the subject. In certain embodiments, the sample comprising DNA, RNA and/or protein is derived from a biopsy sample from the subject. In certain embodiments, the sample comprising DNA, RNA and/or protein is derived from a blood sample. In certain embodiments, the sample comprising DNA, RNA and/or protein is derived from a urine sample.
[00574] Several methods described herein comprise processing the sample, including e.g., processing (e.g., extracting/purifying) DNA, RNA, or protein from the sample.
[00575] Methods for extracting/purifying DNA and/or RNA from a cell, are known in the art. See, e.g., Tan, Siun Chee, and Beow Chin Yiap. “DNA, RNA, and protein extraction: the past and the present.” Journal of biomedicine & biotechnology vol. 2009 (2009): 574398. doi: 10.1155/2009/574398; Ali, Nasir et al. “Current Nucleic Acid Extraction Methods and Their Implications to Point-of-Care Diagnostics.” BioMed research international vol. 2017 (2017): 9306564. doi: 10.1155/2017/9306564; Dou, Yanmei et al. “Detecting Somatic Mutations in Normal Cells.” Trends in genetics: TIG vol. 34,7 (2018): 545-557. doi:10.1016/j.tig.2018.04.003; the entire contents of each of which are incorporated herein by reference for all purposes. For example, DNA and RNA can be isolated from a cell or tissue sample using reagents to lyse cells followed by a column-based approach and/or a bead-based approach to purify the DNA and/or RNA. Examples of commercially available kits that may be used to isolate DNA or RNA from a cell or tissue sample include, but are not limited to the QIAamp® DNA Kit, the EZ1® DNA Tissue Kit, the Oligotex® Direct mRNA Kit, and the Arcturus® PicoPure® DNA Extraction Kit. [00576] The isolated DNA or RNA may be amplified before analysis. Before amplification, the isolated RNA may be converted to DNA by reverse transcription using a reverse transcriptase. DNA or RNA may be amplified using techniques known in the ail, for example by using polymerase chain reaction (PCR)-based methods or reverse transcription polymerase chain reaction (RT- PCR)-based methods. The isolated DNA or RNA may be also be used to construct a DNA or RNA library suitable for the sequencing technique to be used. For example, a DNA library may be constructed using a transposase-based method. The DNA or mRNA isolated from a sample may also be quantified using methods known in the art such as Quantitative PCR (qPCR) and Quantitative reverse transcription PCR (RT-qPCR). Thus, in certain embodiments, the method may further comprise a step of amplifying and/or quantifying the amount of DNA or RNA obtained from a sample obtained from a subject.
[00577] A variety of DNA and RNA sequencing procedures are known in the art and may be used to practice the methods disclosed herein (e.g., for determining the presence of one or more somatic mutation). For example, Sanger sequencing, Polony sequencing, 454 pyrosequencing, Combinatorial probe anchor synthesis, SOLiD sequencing, Ion Torrent semiconductor sequencing, DNA nanoball sequencing, Heliscope single molecule sequencing, Single molecule real time (SMRT) sequencing, Nanopore DNA sequencing, Microfluidic Sanger sequencing and Illumina dye sequencing.
[00578] In embodiments wherein the sample comprises DNA, the DNA may be circulating free DNA (cfDNA). In embodiments wherein the biological sample comprises cfDNA, the method may further comprise isolating the cfDNA from the sample. cfDNA can be isolated from the sample using a variety of techniques known in the art. For example, cfDNA can be isolated by a column- based approach and/or a bead-based approach. In some embodiments, cfDNA is isolated by means of a column-based approach, for example using a commercially available kit such as QIAamp® circulating nucleic acid kit. In some embodiments, cfDNA is isolated by means of a bead-based approach, for example an automated cfDNA extraction system using a commercially available kit such as Maxwell RSC ccfDNA Plasma Kit (Promega). The isolated cfDNA may be amplified before analysis. Thus, the method may further comprise amplification of the isolated cfDNA. Techniques suitable for amplifying cfDNA are known in the art and include, but are not limited to, cloning, polymerase chain reaction (PCR), polymerase chain reaction of specific alleles (PASA), polymerase chain ligation, nested polymerase chain reaction, and so forth. [00579] In embodiments wherein the sample comprises protein, the level of a mutated protein (i.e., encoded by a gene comprising one or more somatic mutations) may be measured using any suitable technique known in the ail. See, e.g., Tan, Siun Chee, and Beow Chin Yiap. “DNA, RNA, and protein extraction: the past and the present.” Journal of biomedicine & biotechnology vol. 2009 (2009): 574398. doi: 10.1155/2009/574398, the entire contents of which are incorporated herein by reference for all purposes. The method may comprise isolating the protein from the sample and then assessing the quantity of the mutated protein present. Depending on the method used and the level of accuracy required, a purification step may be canned out. In some circumstances, simple lysis of cells in the sample may be sufficient. The level of mutated protein present may be assessed using one or more techniques selected from enzyme-linked immunosorbent assay (ELISA), Western Blot analysis, and mass spectrometry.
4.13 Exemplary Diseases
[00580] Several methods described herein comprise a diseases, e.g., methods of treating a disease, methods of diagnosing a disease, methods of prognosticating a disease, etc. (see, e.g., §§ 4.15.1, 4.15.2, 4.15.3, 4.15.4). The following provides additional exemplary diseases that may be utilized in any of the methods described herein.
[00581] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma. In some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00582] In specific embodiments, the disease is a gene (e.g., BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00583] In some embodiments, the disease is a BGT1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. In some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00584] In some embodiments, the disease is a CDKN1B associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00585] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus. In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00586] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00587] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
4.14 Exemplary Therapeutic Agents
[00588] Several methods described herein comprise a therapeutic agent (see, e.g., §§ 4.15.1, 4.15.2, 4.15.3, 4.15.4). The following provides additional exemplary therapeutic agents that may be utilized in any of the methods described herein.
[00589] In some embodiments, the therapeutic agent comprises a nucleic acid molecule, small molecule, protein, peptide.
[00590] Therapeutic agents include, for example, anti-inflammatory agents. Anti-inflammatory agents are known in the art. Exemplary anti-inflammatory agents, include, but are not limited to, angiotensin-converting enzyme 2 (ACE-2) inhibitors (e.g., lisinopril, benazepril, captopril, enalapril, fosinopril, moexipril, perindopril, or quinapril), a corticosteroid (e.g., cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, or hydrocortisone) or a non-steroidal anti-inflammatory drug (NSAID) (e.g., ibuprofen, naproxen, diclofenac, celecoxib, mefenamic acid, etoricoxib, or indomethacin). In one embodiment, the antiinflammatory agent is an agent that blocks, inhibits, or reduces inflammation or signaling from an inflammatory signaling pathway. In one embodiment, the anti-inflammatory agent inhibits or reduces the activity of one or more of any of the following: IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL- 7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, interferons (IFNs), e.g., TNF-a, TNF-p, TNF-RI, TNF-RII; CD23, CD30, CD40L, CXCL-1, EGF, G-CSF, GDNF, PDGF-BB, RANTES/CCL5, IKK, NF-kB, TER2, TLR3, TLR4, TL5, TLR6, TLR7, TLR8, TER8, TLR9, and/or any cognate receptors thereof. [00591] In one embodiment, the anti-inflammatory agent is an IL-1 or IL-1 receptor antagonist, such as anakinra (KINIRET®), rilonaccpt, or canakinumab.
[00592] In one embodiment, the anti-inflammatory agent is an IL-6 or IL-6 receptor antagonist, e.g., an anti-IL-6 antibody or an anti-IL-6 receptor antibody, such as tocilizumab (ACTEMRA®), olokizumab, clazakizumab, sarilumab, sirukumab, siltuximab, or ALX-0061.
[00593] In one embodiment, the anti-inflammatory agent is a TNF-a antagonist, e.g., an anti- TNFa antibody, such as infliximab (REMICADE®), golimumab (SIMPONI®), adalimumab (HUMIRA®), certolizumab pegol (CIMZIA®) or etanercept.
[00594] In one embodiment, the anti-inflammatory agent is a corticosteroid. Exemplary corticosteroids include, but are not limited to, cortisone (hydrocortisone, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, ALA-CORT®, HYDROCORT ACETATE®, hydrocortone phosphate LANACORT®, SOLU-CORTEF®), decadron (dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, DEXASONE®, DIODEX®, HEXADROL®, MAXIDEX®), methylprednisolone (6-methylprednisolone, methylprednisolone acetate, methylprednisolone sodium succinate, DURALONE®, MEDRALONE®, MEDROL®, M-PREDNISOL®, SOLU-MEDROL®), prednisolone (DELTA-CORTEF®, ORAPRED®, PEDIAPRED®, PRELONE®), and prednisone (DELTASONE®, LIQUID PRED®, METICORTEN®, ORASONE®)), and bisphosphonates e.g., pamidronate (AREDIA®), and zoledronic acid (ZOMETA®).
[00595] In another embodiment, the anti-inflammatory agent is a non-steroidal antiinflammatory drug (NSAID). Exemplary anti-inflammatory agents e.g., NSAIDs) include, but are not limited to, aspirin, ibuprofen, naproxen, celecoxib, , diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen, ketoprofen, ketorolac, mefenamic acid, meloxicam, nabumetone, oxaprozin, piroxicam, sulindac, and tolmetin. In an embodiment, the anti-inflammatory agent is an immune selective anti-inflammatory derivative (ImSAID).
[00596] In specific embodiments, the therapeutic agent is an anti-cancer agent. Exemplary anticancer agents include, but are not limited to,
[00597] Examples of chemotherapeutic agents used in cancer therapy include, for example, antimetabolites (e.g., folic acid, purine, and pyrimidine derivatives) and alkylating agents (e.g., nitrogen mustards, nitrosoureas, platinum, alkyl sulfonates, hydrazines, triazenes, aziridines, spindle poison, cytotoxic agents, toposimerase inhibitors and others). Exemplary agents include Aclarubicin, Actinomycin, Alitretinon, Altretamine, Aminopterin, Aminolevulinic acid, Amrubicin, Amsacrinc, Anagrclidc, Arsenic trioxidc, Asparaginase, Atrascntan, Bclotccan, Bexarotene, endamustine, Bleomycin, Bortezomib, Busulfan, Camptothecin, Capecitabine, Carboplatin, Carboquone, Carmofur, Carmustine, Celecoxib, Chlorambucil, Chlormethine, Cisplatin, Cladribine, Clofarabine, Crisantaspase, Cyclophosphamide, Cytarabine, Dacarbazine, Dactinomycin, Daunorubicin, Decitabine, Demecolcine, Docetaxel, Doxorubicin. Efaproxiral, Elesclomol, Elsamitrucin, Enocitabine, Epirubicin, Estramustine, Etoglucid, Etoposide, Floxuridine, Fludarabine, Fluorouracil (5FU), Fotemustine, Gemcitabine, Gliadel implants, Hydroxycarbamide, Hydroxyurea, Idarubicin, Ifosfamide, Irinotecan, Irofulven, Ixabepilone, Larotaxel, Leucovorin, Liposomal doxorubicin, Liposomal daunorubicin, Lonidamine, Lomustine, Lucanthone, Mannosulfan, Masoprocol, Melphalan, Mercaptopurine, Mesna, Methotrexate, Methyl aminolevulinate, Mitobronitol, Mitoguazone, Mitotane, Mitomycin, Mitoxantrone, Nedaplatin, Nimustine, Oblimersen, Omacetaxine, Ortataxel, Oxaliplatin, Paclitaxel, Pegaspargase, Pemetrexed, Pentostatin, Pirarubicin, Pixantrone, Plicamycin, Porfimer sodium, Prednimustine, Procarbazine, Raltitrexed, Ranimustine, Rubitecan, Sapacitabine, Semustine, Sitimagene ceradenovec, Strataplatin, Streptozocin, Talaporfin, Tegafur-uracil, Temoporfin, Temozolomide, Teniposide, Tesetaxel, Testolactone, Tetranitrate, Thiotepa, Tiazofurine, Tioguanine, Tipifamib, Topotecan, Trabectedin, Triaziquone, Triethylenemelamine, Triplatin, Tretinoin, Treosulfan, Trofosfamide, Uramustine, Valrubicin, Verteporfin, Vinblastine, Vincristine, Vindesine, Vinflunine, Vinorelbine, Vorinostat, Zorubicin, and other cytostatic or cytotoxic agents described herein.
[00598] Exemplary anti-cancer agents also include targeted therapies (agents specific for the cancer cells, e.g., a deregulated proteins associated with a cancer cell). Small molecule targeted therapy drugs are generally inhibitors of enzymatic domains on mutated, overexpressed, or otherwise critical proteins within the cancer cell. Prominent examples are the tyrosine kinase inhibitors such as Axitinib, Bosutinib, Cediranib, desatinib, erlotinib, imatinib, gefitinib, lapatinib, Lestaurtinib, Nilotinib, Semaxanib, Sorafenib, Sunitinib, and Vandetanib, and also cyclin- depdendent kinase inhibitors such as Alvocidib and Seliciclib.
[00599] Anti-cancer agents also inlcude, monoclonal antibody therapies. Examples include the anti-HER2/neu antibody trastuzumab (Herceptin®), and the anti-CD20 antibody rituximab and Tositumomab. Other exemplary antibodies include Cetuximab, Panitumumab, Trastuzumab, Alemtuzumab, Bevacizumab, Edrecolomab, and Gemtuzumab. Exemplary fusion proteins include Aflibcrccpt and Dcnilcukin diftitox. Targeted therapy can also involve small peptides as “homing devices” which can bind to cell surface receptors or affected extracellular matrix surrounding the tumor. Radionuclides which are attached to these peptides (e.g., RGDs) eventually kill the cancer cell if the nuclide decays in the vicinity of the cell. An example of such therapy includes BEXXAR®.
[00600] Exemplary anti-cancer therapeutic also include cancer immunotherapies (a diverse set of therapeutic strategies designed to induce the patient's own immune system to fight the tumor). Exemplary immunotherapies include immune checkpoint inhibitors, T-cell therapy, monoclonal antibodies, cancer vaccines, and immune system modulators. Examples of immunotherapies currently under investigation or in use are immune adjuvants, e.g., Mycobacterium bovis, Plasmodium falciparum, dinitrochlorobenzene, and aromatic compounds (U.S. Patents 5,801,005 and 5,739,169; Hui and Hashimoto, 1998; Christodoulides et al, 1998); cytokine therapy, e.g., interferons a, , and y, IL-1, GM-CSF, and TNF (Bukowski et al, 1998; Davidson et al, 1998; Hellstrand et al, 1998); gene therapy, e.g., TNF, IL-1, IL-2, and p53 (Qin et al, 1998; Austin-Ward and Villaseca, 1998; U.S. Patents 5,830,880 and 5,846,945); and monoclonal antibodies, e.g., anti- CD20, anti-ganglioside GM2, and anti-pl85 (Hollander, 2012; Hanibuchi et al, 1998; U.S. Patent 5,824,311). In some embodiments, the immunotherapy may be an immune checkpoint inhibitor. Immune checkpoints either turn up a signal (e.g., co- stimulatory molecules) or turn down a signal. Inhibitory immune checkpoints that may be targeted by immune checkpoint blockade include adenosine A2A receptor (A2AR), B7-H3 (also known as CD276), B and T lymphocyte attenuator (BTLA), cytotoxic T-lymphocyte-associated protein 4 (CTLA-4, also known as CD152), indoleamine 2,3-dioxygenase (IDO), killer-cell immunoglobulin (KIR), lymphocyte activation gene-3 (LAG3), programmed death 1 (PD-1), T-cell immunoglobulin domain and mucin domain 3 (TIM-3) and V-domain Ig suppressor of T cell activation (VISTA). In particular, the immune checkpoint inhibitors target the PD-1 axis and/or CTLA-4. The immune checkpoint inhibitors may be drugs such as small molecules, recombinant forms of ligand or receptors, or, in particular, a e antibodies, such as human antibodies (e.g., International Patent Publication W02015016718; Pardoll, Nat Rev Cancer, 12(4): 252-64, 2012; both incorporated herein by reference). Known inhibitors of the immune checkpoint proteins or analogs thereof may be used, in particular chimerized, humanized or human forms of antibodies may be used. As the skilled person will know, alternative and/or equivalent names may be in use for certain antibodies mentioned in the present disclosure. Such alternative and/or equivalent names arc interchangeable in the context of the present disclosure. For example it is known that lambrolizumab is also known under the alternative and equivalent names MK-3475 and pembrolizumab.
[00601] Additional exemplary therapeutic agents, include, agents targeting GD2 ganglioside (see, e.g., WO 2024049971, the entire contents of which are incorporated herein by reference for all purposes); such an agent may be particularly useful within the context of methods comprising the detection of a BTG1 somatic mutation.
[00602] Additional exemplary therapeutic agents, include, CDK1B degradation inhibitors (see, e.g., WO2014062811, the entire contents of which are incorporated herein by reference for all purposes); (e.g., N-[4-[Bis[4-(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-l-ylidene]- N-methylmethanaminium chloride; 3-(2-Hydroxyethoxy)-7,8,9, 10-tetrahydro-6H- benzo[c]chromen-6-one; CAS # 432001-69-9; CAS# 428844-86-4; CAS# 329918-37-8; such an agent may be particularly useful within the context of methods comprising the detection of a CDK1B somatic mutation.
[00603] In specific embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is a nucleic acid molecule. In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule.
[00604] In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or carrier comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00605] Exemplary agents targeting CDK1B include, e.g., linichlorin A and methylrosaniline chloride. 4.15 Methods of Use
[00606] Provided herein are various methods of utilizing any one or more agent described herein (e.g., and RNA agent (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand)) (see, e.g., §§ 4.2, 4.3); a vector described herein (see, e.g., § 4.7); a conjugate described herein (see, e.g., § 4.4); a carrier described herein (see, e.g., § 4.9); a host cell described herein (see, e.g., § 4.8); and/or a pharmaceutical composition described herein (see, e.g., § 4.10); or any combination thereof.
[00607] In some aspects, the methods described herein comprise administering one or more of the foregoing to a subject. Exemplary subjects include mammals, e.g., humans, non-human mammals, e.g., non-human primates. In some embodiments, the subject is a human.
4.15.1 Methods of Treating, Ameliorating, or Preventing a Disease in a Subject
[00608] Provided herein arc methods of treating, ameliorating, or preventing a disease in a subject, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and (c) administering to the subject a therapeutic agent if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is detected in the DNA, RNA, or protein.
[00609] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA. In some embodiments, (a) comprises isolating and purifying RNA, or having RNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the RNA. In some embodiments, (a) comprises isolating and purifying protein, or having protein isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the protein.
[00610] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
[00611] In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN IB mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00612] In certain embodiments, the therapeutic agent is a therapeutic agent described herein (see, e.g., § 4.14). In specific embodiments, the therapeutic agent is an anti-inflammatory agent (e.g., described herein (see, e.g., § 4.14)). In specific embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is a nucleic acid molecule. In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or earner comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00613] In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for the disease. In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for the disease that is not an inhibitory nucleic acid molecule that inhibits expression of the gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) and/or function of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation in the sample is detected. In some embodiments, the treatment with the different therapeutic agent is discontinued if the therapeutic agent is administered to the subject. In some embodiments, the treatment with the different therapeutic agent is discontinued if the inhibitory nucleic acid molecule that inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) is administered to the subject.
[00614] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma. In some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00615] In specific embodiments, the disease is a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00616] In some embodiments, (i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation in the DNA, RNA, or protein.
[00617] In some embodiments, the disease is a BGT1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. In some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00618] In some embodiments, the disease is a CDKN1B associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00619] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus. In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00620] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00621] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00622] Provided herein are methods of treating, ameliorating, or preventing a disease in a subject, the method comprising administering to the subject any one or more agent described herein, RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein, to thereby treat, ameliorate, or prevent a disease in the subject. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to treat, ameliorate, or prevent the disease in the subject.
[00623] Provided herein are agents, RNA agents, conjugates, vectors, carriers, and pharmaceutical compositions for use in a method of treating, ameliorating, or preventing a disease in a subject, the method comprising administering to the subject an agent described herein, an RNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a pharmaceutical composition described herein, to thereby treat, ameliorate, or prevent the disease in the subject.
[00624] Provided herein are uses of agents, RNA agents, conjugates, vectors, carriers, and pharmaceutical compositions for the manufacture of a medicament for use in a method of treating, ameliorating, or preventing a disease in a subject, the method comprising administering to the subject an agent described herein, an RNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a pharmaceutical composition described herein, to thereby treat, ameliorate, or prevent the disease in the subject.
[00625] Provided herein are agents, RNA agents, conjugates, vectors, carriers, and pharmaceutical compositions for use in treating a disease in a subject, preferably a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease.
[00626] Provided herein are agents, RNA agents, conjugates, vectors, carriers, and pharmaceutical compositions for use in treating a disease in a subject (preferably a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease) by reducing or inhibiting expression of BTG1 (e.g., hBTGl) in a cell.
[00627] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma. In some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00628] In specific embodiments, the disease is a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00629] In some embodiments, the disease is a BGT1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. In some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00630] In some embodiments, the disease is a CD KN IB associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00631] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus. In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00632] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00633] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer {e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer {e.g., a lung adenocarcinoma).
4.15.2 Methods of Diagnosing and/or Prognosticating a Disease in a Subject
[00634] Provided herein are methods of diagnosing a disease in a subject, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified {e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene {e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation {e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the presence of the somatic mutation indicates that the subject has the disease.
[00635] Provided herein are methods of diagnosing and/or prognosticating a disease in a subject, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified {e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene {e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation {e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the presence of the somatic mutation indicates that the subject has a disease, is at risk of developing a disease, is at risk of developing a more severe form of a disease, and/or is at risk of developing a disease associated with a disease.
[00636] For the sake of clarity, it should be understood that the following embodiments apply to any of the foregoing methods described above (as if recited directly below each method).
[00637] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene {e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation {e.g., a somatic mutation set forth in Table 2) in the DNA. In some embodiments, (a) comprises isolating and purifying RNA, or having RNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene {e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation {e.g., a somatic mutation set forth in Table 2) in the RNA. In some embodiments, (a) comprises isolating and purifying protein, or having protein isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the protein.
[00638] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
[00639] In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN IB mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00640] In some embodiments, the method further comprises (c) administering to the subject a therapeutic agent if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
[00641] In some embodiments, the method further comprises (c) administering to the subject a therapeutic agent if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or carrier comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00642] In some embodiments, the method further comprises (c) withholding administration of a therapeutic agent to the subject if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (c) administering an alternative therapeutic agent for treatment of the disease to the subject if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is not detected. In some embodiments, the different therapeutic agent is a standard of care agent for the disease.
[00643] In some embodiments, the method further comprises (c) withholding administration of a therapeutic agent to the subject if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is not detected in the DNA, RNA, or protein, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the method further comprises (c) administering an alternative therapeutic agent for treatment of the disease that is not an inhibitory nucleic acid molecule that inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) to the subject if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is not detected. In some embodiments, the different therapeutic agent is a standard of care agent for the disease.
[00644] In certain embodiments, the therapeutic agent is a therapeutic agent described herein (see, e.g., § 4.14). In specific embodiments, the therapeutic agent is an anti-inflammatory agent (e.g., described herein (see, e.g., § 4.14)). In specific embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is a nucleic acid molecule. In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or earner comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00645] In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for the disease. In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for the disease that is not an inhibitory nucleic acid molecule that inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) and/or function of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation in the sample is detected. In some embodiments, the treatment with the different therapeutic agent is discontinued if the therapeutic agent is administered to the subject. In some embodiments, the treatment with the different therapeutic agent is discontinued if the inhibitory nucleic acid molecule that inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) is administered to the subject.
[00646] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma. In some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00647] In specific embodiments, the disease is a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00648] In some embodiments, (i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation in the DNA, RNA, or protein.
[00649] In some embodiments, the disease is a BGT1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. In some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00650] In some embodiments, the disease is a CD KN IB associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00651] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus . In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00652] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00653] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
4.15.3 Methods of Screening, Identifying, and Selecting a Subject for Treatment with a Therapeutic Agent
[00654] Provided herein are methods of screening a subject for administration of a therapeutic agent, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the subject is selected for administration of the therapeutic agent if the somatic mutation is present.
[00655] Provided herein arc methods of selecting a subject for administration of a therapeutic agent, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the subject is selected for administration of the therapeutic agent if the somatic mutation is present.
[00656] Provided herein are methods of identifying a subject for administration of a therapeutic agent, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the presence of the somatic mutation indicates that the subject is identified for administration of the therapeutic agent.
[00657] Provided herein are methods of identifying a subject having a disease who is likely to respond to treatment with a therapeutic agent, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the subject is identified as a subject likely to respond to treatment with the therapeutic agent if the somatic mutation is present.
[00658] For the sake of clarity, it should be understood that the following embodiments apply to any of the foregoing methods described above (as if recited directly below each method).
[00659] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA. In some embodiments, (a) comprises isolating and purifying RNA, or having RNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the RNA. In some embodiments, (a) comprises isolating and purifying protein, or having protein isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the protein.
[00660] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
[00661] In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00662] In certain embodiments, the therapeutic agent is a therapeutic agent described herein (see, e.g., § 4.14). In specific embodiments, the therapeutic agent is an anti-inflammatory agent (e.g., described herein (see, e.g., § 4.14)). In specific embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is a nucleic acid molecule. In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or carrier comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00663] In some embodiments, the method further comprises (c) administering to the subject the therapeutic agent if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
[00664] In some embodiments, the method further comprises (c) administering to the subject a therapeutic agent, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is detected in the DNA, RNA, or protein. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or carrier comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00665] In some embodiments, the method further comprises (c) withholding administration of the therapeutic agent to the subject if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (c) withholding administration of the therapeutic agent, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) to the subject if one or more somatic gene (e.g., BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6) mutation is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (c) administering an alternative therapeutic agent for treatment of a disease that is not the therapeutic agent (e.g., not an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6)) to the subject if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation in the sample is not detected. In some embodiments, the different therapeutic agent is a standard of care agent for a disease.
[00666] In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for a disease. In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is detected and/or the therapeutic agent is administered to the subject. In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for a disease that is not an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) and/or function of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is detected and/or the inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) is administered to the subject.
[00667] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma. In some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00668] In specific embodiments, the disease is a gene (e.g., BTGI, CDKN IB, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene (e.g., BTGI, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene (e.g., BTGI, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTGI, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00669] In some embodiments, (i) the gene associated disease is a BTGI associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Tabic 2) in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00670] In some embodiments, the disease is a BGT 1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. In some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00671] In some embodiments, the disease is a CDKN1B associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00672] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus. In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00673] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00674] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00675] In specific embodiments, the subject has been diagnosed with one (or more) of the any of the foregoing diseases.
[00676] Provided herein are methods of selecting a therapy for a subject having a disease who is likely to respond to treatment with a therapeutic agent, the method comprising (a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; (b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and (c) selecting a therapy for a subject comprising a therapeutic agent if the somatic mutation is present.
[00677] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA. In some embodiments, (a) comprises isolating and purifying RNA, or having RNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the RNA. In some embodiments, (a) comprises isolating and purifying protein, or having protein isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the protein.
[00678] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
[00679] In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00680] In certain embodiments, the therapeutic agent is a therapeutic agent described herein (see, e.g., § 4.14). In specific embodiments, the therapeutic agent is an anti-inflammatory agent (e.g., described herein (see, e.g., § 4.14)). In specific embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is a nucleic acid molecule. In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or earner comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00681] In some embodiments, the method further comprises (c) administering to the subject a therapeutic agent (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (c) administering to the subject a therapeutic agent, wherein the therapeutic agent inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (c) administering to the subject a therapeutic agent, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is detected in the DNA, RNA, or protein. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or carrier comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00682] In some embodiments, the method further comprises (d) withholding administration of the therapeutic agent to the subject if one or more somatic gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (d) withholding administration of the therapeutic agent, wherein the therapeutic agent inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) to the subject if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (d) withholding administration of the therapeutic agent, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) to the subject if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (d) administering an alternative therapeutic agent for treatment of a disease that is not the therapeutic agent (e.g., not an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6)) to the subject if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is not detected. In some embodiments, the different therapeutic agent is a standard of care agent for a disease.
[00683] In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for a disease. In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is detected and/or the therapeutic agent is administered to the subject. In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for a disease that is not an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) and/or function of gene (e.g., BTGI, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTGI, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is detected and/or the inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTGI, CDKN1B, DUSP2, IGLL5, or KLHL6) is administered to the subject.
[00684] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma, hi some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00685] In specific embodiments, the disease is a gene (e.g., BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00686] In some embodiments, (i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00687] In some embodiments, the disease is a BGT1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. Tn some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00688] In some embodiments, the disease is a CDKN1B associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00689] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus. In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00690] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00691] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00692] In specific embodiments, the subject has been diagnosed with one (or more) of the any of the foregoing diseases.
4.15.4 In Vitro Methods of Screening Samples for Somatic Gene Mutations
[00693] In one aspect, provided herein are methods of screening a sample from a subject for the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2), the method comprising (a) isolating and purifying DNA, RNA, or protein from a sample obtained from the subject; (b) detecting the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00694] In some embodiments, (a) comprises isolating and purifying DNA, or having DNA isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA. In some embodiments, (a) comprises isolating and purifying RNA, or having RNA isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the RNA. In some embodiments, (a) comprises isolating and purifying protein, or having protein isolated and purified, from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the protein.
[00695] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6. [00696] In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein. In some embodiments, (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00697] In certain embodiments, the therapeutic agent is a therapeutic agent described herein (see, e.g., § 4.14). In specific embodiments, the therapeutic agent is an anti-inflammatory agent (e.g., described herein (see, e.g., § 4.14)). In specific embodiments, the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the therapeutic agent is a nucleic acid molecule. In some embodiments, the therapeutic agent is an inhibitory nucleic acid molecule. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a carrier described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or earner comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00698] In some embodiments, the method further comprises (c) administering to the subject the therapeutic agent if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is detected in the DNA, RNA, or protein.
[00699] In some embodiments, the method further comprises (c) administering to the subject a therapeutic agent, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is detected in the DNA, RNA, or protein. In some embodiments, the inhibitory nucleic acid molecule comprises one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand, an siRNA agent, an antisense oligonucleotide) (or a vector comprising or encoding the RNA agent described herein, e.g., a vector described herein; a conjugate comprising the RNA agent described herein, e.g., a conjugate described herein; a carrier comprising the RNA agent described herein (or a vector or conjugate comprising the same), e.g., a earner described herein; and/or a pharmaceutical composition comprising the RNA agent described herein (or a vector, conjugate, or carrier comprising the same), e.g., a pharmaceutical composition described herein). In some embodiments, the RNA agent is a dsRNA agent described herein comprising a sense strand and an antisense strand.
[00700] In some embodiments, the method further comprises (c) withholding administration of the therapeutic agent to the subject if one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (c) withholding administration of the therapeutic agent, wherein the therapeutic agent is an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) to the subject if one or more somatic gene (e.g., BTG1, CD KN IB, DUSP2, IGLL5, or KLHL6) mutation is not detected in the DNA, RNA, or protein. In some embodiments, the method further comprises (c) administering an alternative therapeutic agent for treatment of a disease that is not the therapeutic agent (e.g., not an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6)) to the subject if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation in the sample is not detected. In some embodiments, the different therapeutic agent is a standard of care agent for a disease.
[00701] In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for a disease. In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is detected and/or the therapeutic agent is administered to the subject. In some embodiments, the subject is undergoing or has undergone treatment with a different therapeutic agent for a disease that is not an inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1 , CDKN1B, DUSP2, IGLL5, or KLHL6) and/or function of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6). In some embodiments, the treatment with the different therapeutic agent is discontinued if a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the sample is detected and/or the inhibitory nucleic acid molecule that inhibits expression of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) is administered to the subject.
[00702] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma. In some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00703] In specific embodiments, the disease is a gene (e.g., BTG1, CDKN IB, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00704] In some embodiments, (i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00705] In some embodiments, the disease is a BGT1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. In some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00706] In some embodiments, the disease is a CDKN1B associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00707] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus. In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00708] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00709] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00710] In specific embodiments, the subject has been diagnosed with one (or more) of the any of the foregoing diseases.
[00711] Provided herein are methods of characterizing a DNA molecule in a sample from a subject having a disease, the method comprising (a) isolating and purifying DNA (or having DNA isolated and purified) in a sample from the subject; (b) analyzing the DNA (e.g., sequencing at least a portion of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) gene in the DNA) (e.g., from a cell or population of cells); and (c) determining the presence or absence of a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation in the DNA molecule.
[00712] Provided herein are methods of characterizing a disease in a subject, the method comprising (a) isolating and purifying DNA (or having DNA isolated and purified) in a sample from the subject; (b) detecting the presence or absence of a somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation in the DNA (e.g., from a cell or population of cells); and (c) characterizing the disease as somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation positive or somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation negative based on the detection.
[00713] For the sake of clarity, it should be understood that the following embodiments apply to any of the foregoing methods described above (as if recited directly below each method).
[00714] In some embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6. [00715] In specific embodiments, the disease is a proinflammatory disease. In some embodiments, the disease is an autoimmune disease. In specific embodiments, the disease is inflammatory bowel disease, ulcerative colitis, rheumatoid arthritis, or systemic lupus erythematosus. In some embodiments, the disease is a cancer. In specific embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is a colon adenocarcinoma. In some embodiments, the cancer is a lung cancer. In some embodiments, the cancer is a lung adenocarcinoma. In some embodiments, the disease is a brain disease. In some embodiments, the disease is a brain disease is schizophrenia.
[00716] In specific embodiments, the disease is a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease. In some embodiments, the gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory disease. In some embodiments, the gene e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease. In some embodiments, the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a cancer.
[00717] In some embodiments, (i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, (iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, or (v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
[00718] In some embodiments, the disease is a BGT1 associated disease. In some embodiments, the BTG1 associated disease is a proinflammatory disease. In some embodiments, the BTG1 associated disease is an autoimmune disease. In some embodiments, the BTG1 associated disease is inflammatory bowel disease. In some embodiments, the BTG1 associated disease is ulcerative colitis. In some embodiments, the BTG1 associated disease is schizophrenia.
[00719] In some embodiments, the disease is a CD KN IB associated disease. In some embodiments, the CDKN1B associated disease is a proinflammatory disease. In some embodiments, the CDKN1B associated disease is an autoimmune disease. In some embodiments, the CDKN1B associated disease is systemic lupus erythematosus. In some embodiments, the CDKN1B associated disease is rheumatoid arthritis. In some embodiments, the CDKN1B associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00720] In some embodiments, the disease is a DUSP2 associated disease. In some embodiments, the DUSP2 associated disease is a proinflammatory disease. In some embodiments, the DUSP2 associated disease is an autoimmune disease. In some embodiments, the DUSP2 associated disease is inflammatory bowel disease. In some embodiments, the DUSP2 associated disease is systemic lupus erythematosus. In some embodiments, the DUSP2 associated disease is rheumatoid arthritis. In some embodiments, the DUSP2 associated disease is ulcerative colitis. In some embodiments, the DUSP2 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma). [00721] In some embodiments, the disease is a IGLL5 associated disease. In some embodiments, the IGLL5 associated disease is a proinflammatory disease. In some embodiments, the IGLL5 associated disease is an autoimmune disease. In some embodiments, the IGLL5 associated disease is rheumatoid arthritis. In some embodiments, the IGLL5 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
[00722] In some embodiments, the disease is a KLHL6 associated disease. In some embodiments, the KLHL6 associated disease is a proinflammatory disease. In some embodiments, the KLHL6 associated disease is an autoimmune disease. In some embodiments, the KLHL6 associated disease is inflammatory bowel disease. In some embodiments, the KLHL6 associated disease is a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is an adenocarcinoma. In some embodiments, the cancer is a colon cancer (e.g., a colon adenocarcinoma). In some embodiments, the cancer is a lung cancer (e.g., a lung adenocarcinoma).
4.15.5 Methods of Delivery
[00723] Provided herein are, inter alia, various methods of delivering any one or more agent described herein (e.g., an RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand)); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein; or any combination thereof to e.g., a cell, subject, a cell within a subject.
[00724] In one aspect, provided herein are methods of delivering any one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein; or any combination thereof to a cell, the method comprising introducing into a cell any one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein into the cell, to thereby deliver the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition into the cell. In some embodiments, the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to deliver the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition into the cell.
[00725] In some embodiments, the cell is in vitro, ex vivo, or in vivo. In some embodiments, the cell is in vitro or ex vivo. In some embodiments, the cell is in vitro. In some embodiments, the cell is ex vivo. In some embodiments, the cell is in vivo. [00726] In some embodiments, the cell is an immune cell. Exemplary immune cells include, T cells (e.g., CD4+ T cells, CD8+ T cells), B-cclls, tumor infiltrating lymphocytes (e.g., tumor infiltrating T cells (e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells)). In some embodiments, the cell is a T-cell. In some embodiments, the cell is a tumor infiltrating lymphocyte. In some embodiments, the cell is a tumor infiltrating T cell. In some embodiments, the cell is a tumor infiltrating CD4+ T cell. In some embodiments, the cell is a CD8+ tumor infiltrating T cell. In some embodiments, the cell is B-cell.
[00727] In one aspect, provided herein are methods of delivering any one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein; or any combination thereof to a subject, the method comprising administering to a subject any one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein into the cell, to thereby deliver the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition to the subject. In some embodiments, the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is administered in an amount and for a time sufficient to deliver the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition to the subject.
[00728] In one aspect, provided herein are methods of delivering any one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein; or any combination thereof to a cell within a subject, the method comprising administering to a cell within a subject any one or more RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein into the cell, to thereby deliver the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition to the cell within the subject. In some embodiments, the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is administered in an amount and for a time sufficient to deliver the RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition to the cell within the subject.
[00729] In some embodiments, the cell is an immune cell. Exemplary immune cells include, T cells (e.g., CD4+ T cells, CD8+ T cells), B-cells, tumor infiltrating lymphocytes (e.g., tumor infiltrating T cells (e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells)). In some embodiments, the cell is a T-cell. In some embodiments, the cell is a tumor infiltrating lymphocyte. In some embodiments, the cell is a tumor infiltrating T cell. In some embodiments, the cell is a tumor infiltrating CD4+ T cell. In some embodiments, the cell is a CD8+ tumor infiltrating T cell. In some embodiments, the cell is B-cell.
4.15.6 Methods of Reducing or Inhibiting Gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) Expression
[00730] In one aspect, provided herein are methods of reducing or inhibiting gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in a cell, the method comprising introducing into the cell any one or more agent described herein (e.g., an RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand)); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein, to thereby reduce or inhibit gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell. [00731] In some embodiments, the cell is in vitro, ex vivo, or in vivo. In some embodiments, the cell is in vitro or ex vivo. In some embodiments, the cell is in vitro. In some embodiments, the cell is ex vivo. In some embodiments, the cell is in vivo.
[00732] In some embodiments, the cell is an immune cell. Exemplary immune cells include, T cells (e.g., CD4+ T cells, CD8+ T cells), B-cells, tumor infiltrating lymphocytes (e.g., tumor infiltrating T cells (e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells)). In some embodiments, the cell is a T-cell. In some embodiments, the cell is a tumor infiltrating lymphocyte. In some embodiments, the cell is a tumor infiltrating T cell. In some embodiments, the cell is a tumor infiltrating CD4+ T cell. In some embodiments, the cell is a CD8+ tumor infiltrating T cell. In some embodiments, the cell is B-ccll.
[00733] In specific embodiments, the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6. In specific embodiments, the gene is BTG1. In specific embodiments, the gene is CDKN1B. In specific embodiments, the gene expressed is DUSP2. In specific embodiments, the gene is IGLL5. In specific embodiments, the gene is KLHL6.
[00734] In specific embodiments, the gene is hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6. In specific embodiments, the gene is hBTGl. In specific embodiments, the gene is hCDKNIB. In specific embodiments, the is hDUSP2. In specific embodiments, the gene expressed is hIGLL5. In specific embodiments, the gene is hKLHL6.
[00735] In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 50% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 75% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 80% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 90% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 95% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR).
[00736] In one aspect, provided herein are methods of reducing or inhibiting expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject, the method comprising administering to a subject any one or more agent described herein, RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject. In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR).
[00737] In one aspect, provided herein are agents, RNA agents (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugates, vectors, carriers, and pharmaceutical compositions for use in a method of reducing or inhibiting expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject, the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject. In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR).
[00738] In one aspect, provided herein are uses of agents, RNA agents, (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugates, vectors, carriers, and pharmaceutical compositions for the manufacture of a medicament for use in a method of reducing or inhibiting expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject, the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject. In some embodiments, the level of gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) expression in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). [00739] In some embodiments, the cell is an immune cell. Exemplary immune cells include, T cells (e.g., CD4+ T cells, CD8+ T cells), B-cells, tumor infiltrating lymphocytes (e.g., tumor infiltrating T cells (e.g., tumor infiltrating CD4+ T cells, tumor infiltrating CD8+ T cells)). In some embodiments, the cell is a T-cell. In some embodiments, the cell is a tumor infiltrating lymphocyte. In some embodiments, the cell is a tumor infiltrating T cell. In some embodiments, the cell is a tumor infiltrating CD4+ T cell. In some embodiments, the cell is a CD8+ tumor infiltrating T cell. In some embodiments, the cell is B-cell.
[00740] In one aspect, provided herein are methods of reducing or inhibiting expression of one or more gene expressed by a population of immune cells (e.g., T cells) in a cell, the method comprising introducing into the cell any one or more agent described herein (e.g., an RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand)); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell. In some embodiments, the cell is in vitro, ex vivo, or in vivo. In some embodiments, the cell is in vitro or ex vivo. In some embodiments, the cell is in vitro. In some embodiments, the cell is ex vivo. In some embodiments, the cell is in vivo. In some embodiments, the method is in vitro. In some embodiments, the cell is a B-cell. In some embodiments, the cell is a T-ccll.
[00741] In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is BTG1. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is CDKN1B. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is DUSP2. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is IGLL5. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is KLHL6.
[00742] In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hBTGl, hCDKNIB, hDUSP2, hIGLLS, or hKLHL6. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hBTGl. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hCDKNIB. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hDUSP2. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hIGLL5. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hKLHL6.
[00743] In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell is reduced by at least about 50% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell is reduced by at least about 75% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) expression in the cell is reduced by at least about 80% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell is reduced by at least about 90% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell is reduced by at least about 95% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR).
[00744] In one aspect, provided herein are methods of reducing or inhibiting expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in a cell in a subject, the method comprising administering to a subject any one or more agent described herein, RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand); a vector described herein; a conjugate described herein; a carrier described herein; a host cell described herein; and/or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell in the subject. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell in the subject. In some embodiments, the level of one or more gene expressed by a subset of immune cells (e.g., T cells) expression in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR).
[00745] In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is BTG1. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is CDKN1B. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is DUSP2. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is IGLL5. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is KLHL6.
[00746] In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hBTGl, hCDKNIB, hDUSP2, hIGLL5, or hKLHL6. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hBTGl. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hCDKNIB. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hDUSP2. Tn specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hIGLL5. In specific embodiments, the gene expressed by a subset of immune cells (e.g., T cells) is hKLHL6.
[00747] In one aspect, provided herein are agents, RNA agents (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugates, vectors, carriers, and pharmaceutical compositions for use in a method of reducing or inhibiting expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in a cell in a subject, the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell in the subject. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell in the subject. In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the cell is a B-cell. In some embodiments, the cell is a T-cell.
[00748] In one aspect, provided herein are uses of agents, RNA agents, (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugates, vectors, carriers, and pharmaceutical compositions for the manufacture of a medicament for use in a method of reducing or inhibiting expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in a cell in a subject, the method comprising administering to the subject a dsRNA agent described herein, a conjugate described herein, a vector described herein, a carrier described herein, or a pharmaceutical composition described herein, to thereby reduce or inhibit expression of one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell in the subject. In some embodiments, the agent, RNA agent (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), conjugate, vector, carrier, or pharmaceutical composition is introduced in an amount and for a time sufficient to reduce or inhibit expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell in the subject. In some embodiments, the level of expression of the one or more gene expressed by a subset of immune cells (e.g., T cells) in the cell is reduced by at least about 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% (e.g., as compared to suitable control) (e.g., as measured by an assay known in the art or described herein, e.g., qPCR). In some embodiments, the cell is a B-cell. In some embodiments, the cell is a T-cell.
4.16 Kits
[00749] In a one aspect, provided herein are kits comprising any one or more agent described herein, RNA agent described herein (e.g., an RNAi agent, a dsRNA agent, an antisense strand, a sense strand) (see, e.g., §§ 4.2, 4.3); a vector described herein (see, e.g., § 4.7); a conjugate described herein (see, e.g., § 4.4); a carrier described herein (see, e.g., § 4.9); a host cell described herein (see, e.g., § 4.8); and/or a pharmaceutical composition described herein (see, e.g., § 4.10); or any combination thereof. In addition, the kit may comprise a liquid vehicle for solubilizing or diluting, and/or technical instructions. The technical instructions of the kit may contain information about administration and dosage and subject groups.
[00750] In some embodiments, the agent (e.g., described herein), RNA agent e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), vector (e.g., described herein), conjugate (e.g., described herein), carrier (e.g., described herein), host cell (e.g., described herein), and/or pharmaceutical composition (e.g., described herein) is provided in a separate part of the kit. In some embodiments, the agent (e.g., described herein), RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), vector (e.g., described herein), conjugate (e.g., described herein), carrier (e.g., described herein), host cell (e.g., described herein), and/or pharmaceutical composition (e.g., described herein) is optionally lyophilized, spray-dried, or spray-freeze dried. The kit may further contain as a part a vehicle (e.g., buffer solution) for solubilizing the dried or lyophilized any agent (e.g., described herein), RNA agent (e.g., described herein) (e.g., RNAi agent, dsRNA agent, antisense strand, sense strand), vector (e.g., described herein), conjugate (e.g., described herein), carrier (e.g., described herein), host cell (e.g., described herein), and/or pharmaceutical composition (e.g., described herein).
[00751] In some embodiments, the kit comprises a single dose container. In some embodiments, the kit comprises a multi-dose container. In some embodiments, the kit comprises an administration device (e.g., an injector for intradermal injection or a syringe for intramuscular injection).
[00752] In one aspect, provided herein arc in vitro test kits for use in a method described herein (e.g., diagnostic method, prognostic method, subject screening, subject identification, subject selection, sample screening). In some embodiments, the kit comprises one or more reagents for detecting one or more somatic mutation (e.g., in BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a sample from a subject (e.g., in DNA, RNA, or protein from a sample from a subject).
[00753] Any of the kits described herein may be used in any of the methods described herein (see, e.g., § 4.15).
5. EXAMPLES
TABLE OF CONTENTS
5.1 Example 1. BTG1 Expression in Inflammatory Bowel Disease.
5.2 Example 2. CDKN1B Expression in Cancer.
5.3 Example 3. DUSP2 Expression in Cancer and Inflammatory Bowel Disease.
5.4 Example 4. IGLL5 Expression in Cancer.
5.5 Example 5. KLHL6 Expression in Cancer and Inflammatory Bowel Disease.
5.1 Example 1. BTG1 Expression in Inflammatory Bowel Disease.
[00754] 20 tissue samples (B-cell extracts) from 20 subjects with inflammatory bowel disease
(IBD) were sequenced using NanoSeq technology to a total depth of 3450 duplex coverage. Somatic nucleotide mutations were called using standard bioinformatics algorithms and analysed using the expected to observed ratio of nonsynonymous to synonymous SNPs. Results of interest in the sample were exome-wide defined as a global q-value< 0.1.
[00755] BTG1 was found to be under significant selection in B-cells in IBD with selection parameter estimate of 11.3, p-value of 1.796226e-07 and q-value of 3.41e-04. The selection signal is driven through missense mutations (n=l 1 mutations in 5 donors), suggesting gain of function as the potential mechanism of action.
5.2 Example 2. CDKN1B Expression in Cancer.
[00756] 90 tissue samples (45 CD4+ T-cell extracts and 45 CD8+ T-cell extracts) from 45 subjects with lung adenocarcinoma or colon adenocarcinoma patients were sequenced using NanoSeq technology to a total depth of 22,450 duplex coverage. Somatic nucleotide mutations were called using standard bioinformatics algorithms and analysed using the expected to observed ratio of nonsynonymous to synonymous SNPs. Results of interest in the sample were exome-wide defined as a global q value<0.1.
[00757] CDKN1B was found to be under significant selection in tumour infiltrating lymphocytes with selection parameter estimate of 29.7, p-value of 3.40E-08 and q-value of 1.70E- 04 for CD4+ T-cells, and selection parameter estimate of 22.3, p-value of 2.7e-07 and q-value of 1. 2.7e-03 for CD8+ T-cells. The selection signal is driven through truncating mutations and INDELs (17 mutations in 9 donors), suggesting loss of function as the potential mechanism of action.
5.3 Example 3. DUSP2 Expression in Cancer and Inflammatory Bowel Disease.
[00758] 20 tissue samples (B-cell extracts) from 20 subjects with inflammatory bowel disease
(IBD), and 19 tissue samples (B-cell extracts) from 19 subjects with lung adenocarcinoma or colon adenocarcinoma were sequenced using NanoSeq technology to a total depth of 3450 and 1462 duplex coverage, respectively. Somatic nucleotide mutations were called using standard bioinformatics algorithms and analysed using the expected to observed ratio of nonsynonymous to synonymous SNPs. Results of interest in the sample were exome-wide defined as a global q- value < 0.1 .
[00759] DUSP2 was found to be under significant selection in tumor infiltrating B-cells with selection parameter estimate of 22.2, p-value of 1.10E-06 and q-value of 3.00E-03, and in B-cells in IBD with selection parameter estimate of 16.72, p-value of <2.2e-16 and q-value of <2.2e-16. The selection signal is driven through missense mutations (8 mutations in 6 donors in tumour infiltrating B-cells, and 25 mutations in 14 donors in IBD), suggesting gain of function as the potential mechanism of action.
5.4 Example 4. IGLL5 Expression in Cancer.
[00760] 19 tissue samples (B-cell extracts) from 19 subjects with lung adenocarcinoma or colon adenocarcinoma were sequenced using NanoSeq technology to a total depth of 1462 duplex coverage. Somatic nucleotide mutations were called using standard bioinformatics algorithms and analysed using the expected to observed ratio of nonsynonymous to synonymous SNPs. Results of interest in the sample were exome-wide defined as a global q-value < 0.1.
[00761] IGLL5 was found to be under significant selection in tumor infiltrating B -cells with selection parameter estimate of 21.4, p-value of 3.90E-61 and q-value of 7.70E-57. The selection signal is driven through missense mutations (76 mutations in 16 donors).
5.5 Example 5. KLHL6 Expression in Cancer and Inflammatory Bowel Disease.
[00762] 20 tissue samples (B-cell extracts) from 20 subjects with inflammatory bowel disease
(IBD), and 19 tissue samples (B-cell extracts) from 19 subjects with lung adenocarcinoma or colon adenocarcinoma were sequenced using NanoSeq technology to a total depth of 3450 and 1462 duplex coverage, respectively. Somatic nucleotide mutations were called using standard bioinformatics algorithms and analysed using the expected to observed ratio of nonsynonymous to synonymous SNPs. Results of interest in the sample were exome-wide defined as a global q- value < 0.1.
[00763] KLHL6 was found to be under significant selection in tumor infiltrating B-cells with selection parameter estimate of 13.9, p-value of 4.20E-05 and q-value of 7.00E-02 and in B-cells in IBD with selection parameter estimate of 27.93, p-value of <2.2e-16 and q-value of <2.2e-16. The selection signal is driven through missense mutation (9 mutations in 7 donors in tumour infiltrating B-cells, and 30 mutations in 9 donors in IBD) suggesting gain of function as the potential mechanism of action.
* * *
[00764] The invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.
[00765] All references (e.g., publications or patents or patent applications) cited herein are incorporated herein by reference in their entireties and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes.
[00766] Other embodiments are within the following claims.

Claims

CLAIMS What is claimed is:
1. A method of treating, ameliorating, or preventing a CDKN1B, BTG1, DUSP2, IGLL5, or KLHL6 associated disease in a subject, the method comprising
(a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject;
(b) detecting, or having detected, the presence or absence of one or more somatic CDKN1B, BTG1, DUSP2, IGLL5, or KLHL6 mutation set forth in Table 2 in the DNA, RNA, or protein, and
(c) administering to the subject a therapeutic agent if the one or more somatic CDKN1B, BTG1, DUSP2, IGLL5, or KLHL6 mutation set forth in Table 2 is detected in the DNA, RNA, or protein.
2. The method of claim 1, wherein the gene is CDKN IB and the one or more somatic mutation and the one or more somatic mutation comprises one or more of chrl2:12717949:A- >T; chrl2:12718008:C->T; chrl2:12717909:CC->TG; chrl2:12717940:GC->G; or chrl2:12718016:G->C.
3. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of BTG1 (e.g., human BTG1 (hBTGl)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
4. The dsRNA agent of claim 3, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1.
5. The dsRNA agent of claim 3 or 4, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
6. A dsRNA agent for inhibiting expression of BTG1 (e.g., (hBTGl) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding BTG1 (e.g., (hBTGl).
7. The dsRNA agent of claim 6, wherein the region of complementarity is fully complementary to the portion of the mRNA sequence encoding BTG1 (e.g., (hBTGl).
8. The dsRNA agent of claim 6 or 7, wherein the region of complementarity comprises at least 15 contiguous nucleotides.
9. The dsRNA agent of any one of claims 6-8, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
10. The dsRNA agent of any one of claims 6-9, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1.
11. The dsRNA agent of any one of claims 6-10, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
12. A dsRNA agent for inhibiting expression of BTG1 (e.g., (hBTGl) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding BTG1 (e.g., (hBTGl), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 2.
13. The dsRNA agent of claim 12, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 1.
14. The dsRNA agent of claim 12 or 13, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 1 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 2.
15. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of CDKN1B (e.g., human CDKN1B (hCDKNIB)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
16. The dsRNA agent of claim 15, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4.
17. The dsRNA agent of claim 15 or 16, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
18. A dsRNA agent for inhibiting expression of CDKN1B (e.g., (hCDKNIB) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding CDKN1B (e.g., (hCDKNIB).
19. The dsRNA agent of claim 18, wherein the region of complementarity is fully complementary to the portion of the mRNA sequence encoding CDKN1B (e.g., (hCDKNIB).
20. The dsRNA agent of claim 18 or 19, wherein the region of complementarity comprises at least 15 contiguous nucleotides.
21. The dsRNA agent of any one of claims 18-20, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
22. The dsRNA agent of any one of claims 18-21, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4.
23. The dsRNA agent of any one of claims 18-22, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
24. A dsRNA agent for inhibiting expression of CDKN1B (e.g., (hCDKNIB) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding CDKN1B (e.g., (hCDKNIB), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 5.
25. The dsRNA agent of claim 24, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 4.
26. The dsRNA agent of claim 24 or 25, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 4 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 5.
27. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of DUSP2 (e.g., human DUSP2 (hDUSP2)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
28. The dsRNA agent of claim 27, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7.
29. The dsRNA agent of claim 27 or 28, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
30. A dsRNA agent for inhibiting expression of DUSP2 (e.g., (hDUSP2) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding DUSP2 (e.g., (hDUSP2).
31. The dsRNA agent of claim 30, wherein the region of complementarity is fully complementary to the portion of the mRNA sequence encoding DUSP2 (e.g., (hDUSP2).
32. The dsRNA agent of claim 30 or 31, wherein the region of complementarity comprises at least 15 contiguous nucleotides.
33. The dsRNA agent of any one of claims 30-32, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
34. The dsRNA agent of any one of claims 30-33, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7.
35. The dsRNA agent of any one of claims 30-34, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
36. A dsRNA agent for inhibiting expression of DUSP2 (e.g., (hDUSP2) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding DUSP2 (e.g., (hDUSP2), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 8.
37. The dsRNA agent of claim 36, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 7.
38. The dsRNA agent of claim 36 or 37, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 7 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 8.
39. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of IGLL5 (e.g., human IGLL5 (hIGLLS)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
40. The dsRNA agent of claim 39, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10.
41. The dsRNA agent of claim 39 or 40, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
42. A dsRNA agent for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding IGLL5 (e.g., (hIGLL5).
43. The dsRNA agent of claim 42, wherein the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 (e.g., (hIGLL5).
44. The dsRNA agent of claim 42 or 43, wherein the region of complementarity comprises at least 15 contiguous nucleotides.
45. The dsRNA agent of any one of claims 42-44, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
46. The dsRNA agent of any one of claims 42-45, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10.
47. The dsRNA agent of any one of claims 42-46, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
48. A dsRNA agent for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 11.
49. The dsRNA agent of claim 48, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 10.
50. The dsRNA agent of claim 48 or 49, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 10 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 11.
51. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of IGLL5 (e.g., human IGLL5 (hIGLL5)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
52. The dsRNA agent of claim 51, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13.
53. The dsRNA agent of claim 51 or 52, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
54. A dsRNA agent for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding IGLL5 (e.g., (hIGLL5).
55. The dsRNA agent of claim 54, wherein the region of complementarity is fully complementary to the portion of the mRNA sequence encoding IGLL5 (e.g., (hIGLL5).
56. The dsRNA agent of claim 54 or 55, wherein the region of complementarity comprises at least 15 contiguous nucleotides.
57. The dsRNA agent of any one of claims 54-56, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
58. The dsRNA agent of any one of claims 54-57, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13.
59. The dsRNA agent of any one of claims 54-58, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
60. A dsRNA agent for inhibiting expression of IGLL5 (e.g., (hIGLL5) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding IGLL5 (e.g., (hIGLL5), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 14.
61. The dsRNA agent of claim 60, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 13.
62. The dsRNA agent of claim 60 or 61, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 13 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 14.
63. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of KLHL6 (e.g., human KLHL6 (hKLHL6)) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
64. The dsRNA agent of claim 63, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16.
65. The dsRNA agent of claim 63 or 64, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
66. A dsRNA agent for inhibiting expression of KLHL6 (e.g., (hKLHL6) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to a portion of an mRNA sequence encoding KLHL6 (e.g., (hKLHL6).
67. The dsRNA agent of claim 66, wherein the region of complementarity is fully complementary to the portion of the mRNA sequence encoding KLHL6 (e.g., (hKLHL6).
68. The dsRNA agent of claim 66 or 67, wherein the region of complementarity comprises at least 15 contiguous nucleotides.
69. The dsRNA agent of any one of claims 66-68, wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
70. The dsRNA agent of any one of claims 66-69, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16.
71. The dsRNA agent of any one of claims 66-70, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
72. A dsRNA agent for inhibiting expression of KLHL6 (e.g., (hKLHL6) comprising a sense strand and an antisense strand that form a double stranded region, wherein the antisense strand comprises a region of complementarity to an mRNA encoding KLHL6 (e.g., (hKLHL6), and wherein the region of complementarity comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4 or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 17.
73. The dsRNA agent of claim 72, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the nucleotide sequence of SEQ ID NO: 16.
74. The dsRNA agent of claim 72 or 73, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 5 (e.g., 0, 1, 2, 3, 4, or 5) nucleotides from the region of SEQ ID NO: 16 that is complementary to the sequence of the antisense strand from the nucleotide sequence of SEQ ID NO: 17.
75. The dsRNA agent of any one of claims 3-74, wherein the sense strand comprises at least one modified nucleotide and/or the antisense strand comprises at least one modified nucleotide.
76. The dsRNA agent of any one of claims 3-75, wherein at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the sense strand and/or antisense strand are modified.
77. The dsRNA agent of any one of claims 3-76, wherein substantially all (or all) of the nucleotides in the sense strand and/or antisense strand are modified.
78. The dsRNA agent of any one of claims 3-77, wherein at least one of the modified nucleotides comprises a modified sugar- (e.g., ribose moiety).
79. The dsRNA agent of any one of claims 3-78, wherein at least one of the modified nucleotides comprises a modified nucleobase.
80. The dsRNA agent of any one of claims 3-79, wherein the sense strand comprises at least one modified internucleoside linkage and/or the antisense strand comprises at least one modified intemucleoside linkage.
81. The dsRNA agent of any one of claims 3-80, wherein each of the antisense strand and the sense strand are not more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides in length.
82. The dsRNA agent of any one of claims 3-81, wherein the antisense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-3020-30, 21-30, 22-30, 23-30, 24-30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides; and/or the sense strand comprises from about 15-30, 16-30, 17-30, 18-30, 19-30 20-30, 21-30, 22-30, 23-30, 24- 30, 25-30, 36-30, 27-30, 28-30-, 29-30, 19-20, 19-21, 19-22, 19-23, 19-24, or 19-25 nucleotides.
83. The dsRNA agent of any one of claims 3-82, wherein the antisense strand comprises or consists of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides; and/or the antisense strand comprises or consist of about 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides.
84. The dsRNA agent of any one of claims 3-83, wherein antisense strand comprises from about 19-23 nucleotides; and/or the sense strand comprises from about 19-23 nucleotides.
85. The dsRNA agent of any one of claims 3-84, wherein antisense strand comprises or consists of about 23 nucleotides; and/or the sense strand comprises or consists of about 21 nucleotides.
86. The dsRNA agent of any one of claims 3-85, wherein the sense strand and/or the antisense strand comprises a 3' and/or 5' overhang of 1, 2, or 3 nucleotides.
87. The dsRNA agent of any one of claims 3-86, wherein the antisense strand comprises a 3' overhang of 1, 2, or 3 nucleotides 2 nucleotides).
88. The dsRNA agent of any one of claims 3-87, wherein the double stranded region is from about 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-20, 19-21, 23-30, 23- 29, 23-28, 23-27, 23-26, 23-25, 23-24, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotide pairs in length.
89. The dsRNA agent of any one of claims 3-88, wherein the double stranded region is from about 19-23 or 19-21 nucleotide pairs in length.
90. The dsRNA agent of any one of claims 3-89, wherein the double stranded region is about 21 nucleotide pairs in length.
91. The dsRNA agent of any one of claims 3-90, wherein the sense strand and the antisense strand are part of a single nucleic acid molecule (e.g., wherein a hairpin loop is between the sense strand and the antisense strand of the single nucleic acid molecule).
92. The dsRNA agent of any one of claims 3-91, wherein the sense strand and the antisense strand are separate nucleic acid molecules (z.e., connected only through the double stranded region).
93. A conjugate comprising the dsRNA agent of any one of claims 3-92 and a heterologous moiety.
94. The conjugate of claim 93, wherein the heterologous moiety is a peptide, protein, carbohydrate, lipid, polymer, or small molecule.
95. The conjugate of any one of claims 93-94, wherein the heterologous moiety is attached to the dsRNA agent via a linker.
96. The conjugate of any one of claims 93-95, wherein the heterologous moiety attached to the 3 ' end of the sense and/or antisense strand and/or the 5 ' end of the sense and/or antisense strand, and/or at an internal site of the sense and/or antisense strand.
97. A vector (e.g., a viral vector, a non-viral vector) encoding the antisense strand, the sense strand, or both the antisense and sense strand of any one of claims 3-92, the dsRNA agent of any one of claims 3-92, of the conjugate of any one of claims 93-96.
98. A carrier comprising the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, or the vector of claim 97.
99. The carrier of claim 98, wherein the carrier comprises a nanoparticle, a polymer, a lipid- based delivery system, as dendrimer, a cationic delivery system, or a hydrogel.
100. The carrier of claim 99, wherein the lipid-based delivery system is a lipid nanoparticle (LNP), liposome, lipoplex, nanoliposome, an exosome, or a micelle.
101. A cell comprising the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, or the carrier of any one of claims 98-100.
102. A pharmaceutical composition comprising the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, or the cell of claim 101, and a pharmaceutically acceptable excipient.
103. A kit comprising the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102.
104. A diagnostic kit comprising one or more reagent for use in a method of detecting the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation described herein, e.g., a somatic mutation set forth in Table 2 herein) in a sample from a subject.
105. A method of treating, ameliorating, or preventing a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in a subject, the method comprising
(a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject;
(b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and (c) administering to the subject a therapeutic agent if the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
106. The method of claim 105, wherein the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
107. The method of claim 105 or 106, wherein the one or more somatic mutation is one or more somatic mutation set forth herein.
108. The method of any one of claims 105-107, wherein the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
109. The method of any one of claims 105-108, wherein the gene associated disease is a BTG1 associated disease, a CDKN1B associated disease, DUSP2 associated disease, a IGLL5 associated disease, or a KLHL6 associated disease.
110. The method of any one of claims 105-109, wherein
(i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells),
(ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells),
(iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells),
(iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells), or
(v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells).
111. The method of any one of claims 105-110, wherein the therapeutic agent comprises an anti-inflammatory agent e.g., an anti-inflammatory agent described herein).
112. The method of any one of claims 105-111, wherein the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
113. The method of any one of claims 105-112, wherein the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
114. The method of claim 113, wherein the inhibitory nucleic acid molecule comprises the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102.
115. A method of treating, ameliorating, or preventing a proinflammatory (e.g., autoimmune) disease in a subject, the method comprising
(a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject;
(b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, and
(c) administering to the subject a therapeutic agent if the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
116. The method of claim 115, wherein the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
117. The method of claim 115 or 116, wherein the one or more somatic mutation is one or more somatic mutation set forth herein.
118. The method of any one of claims 115-117, wherein the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
119. The method of any one of claims 115-118, wherein the therapeutic agent is an antiinflammatory agent (e.g., described herein).
120. The method of any one of claims 1 15- 119, wherein the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
121. The method of any one of claims 115-120, wherein the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
122. The method of claim 121, wherein the inhibitory nucleic acid molecule comprises the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102.
123. A method of diagnosing a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in a subject, the method comprising
(a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject;
(b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the presence of the one or more somatic mutation (e.g., a somatic mutation set forth in Table 2) indicates that the subject has a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease.
124. The method of claim 123, wherein the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
125. The method of claim 123 or 124, wherein the one or more somatic mutation is one or more somatic mutation set forth herein.
126. The method of any one of claims 123-125, wherein the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
127. The method of any one of claims 123-126, wherein the gene is associated disease is a BTG1 associated disease, a CDKN1B associated disease, DUSP2 associated disease, a IGLL5 associated disease, or a KLHL6 associated disease.
128. The method of any one of claims 123-127, wherein
(i) the gene associated disease is a BTG1 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic BTG1 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells),
(ii) the gene associated disease is a CDKN1B associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic CDKN1B mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells),
(iii) the gene associated disease is a DUSP2 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic DUSP2 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells),
(iv) the gene associated disease is a IGLL5 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic IGLL5 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells), or
(v) the gene associated disease is a KLHL6 associated disease and (b) comprises detecting, or having detected, the presence or absence of one or more somatic KLHL6 mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein (e.g., from a cell or population of cells).
129. The method of any one of claims 123-128, wherein the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is a proinflammatory (e.g., autoimmune) disease.
130. The method of any one of claims 123-129, wherein the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is an autoimmune disease.
131. The method of any one of claims 123-130, wherein (a) comprises isolating and purifying DNA, or having DNA isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
132. The method of any one of claims 123-131, further comprising administering to the subject a therapeutic agent if a somatic mutation (e.g., a somatic mutation set forth in Table 2) is detected in the DNA, RNA, or protein.
133. The method of any one of claims 123- 132, wherein the therapeutic agent is an antiinflammatory agent (e.g., described herein).
134. The method of any one of claims 123-133, wherein the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
135. The method of any one of claims 123-134, wherein the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
136. The method of claim 135, wherein the inhibitory nucleic acid molecule comprises the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102.
137. A method of selecting a subject for administration of a therapeutic agent, the method comprising
(a) isolating and purifying DNA, RNA, or protein, or having DNA, RNA, or protein isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject;
(b) detecting, or having detected, the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein, wherein the subject is selected for administration of the therapeutic agent if the one or more somatic mutation (e.g., a somatic mutation set forth in Table 2) is present.
138. The method of claim 137, wherein (a) comprises isolating and purifying DNA, or having DNA isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
139. The method of claim 137 or 138, wherein the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
140. The method of any one of claims 137-139, wherein the one or more somatic mutation is one or more somatic mutation set forth herein.
141. The method of any one of claims 137-140, wherein the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
142. The method of any one of claims 137- 141 , further comprising administering to the subject the therapeutic agent if the subject is selected.
143. The method of any one of claims 137-142, wherein the therapeutic agent is an antiinflammatory agent (e.g., described herein).
144. The method of any one of claims 137-143, wherein the therapeutic agent inhibits expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
145. The method of any one of claims 137-144, wherein the therapeutic agent is an inhibitory nucleic acid molecule that targets the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
146. The method of claim 145, wherein the inhibitory nucleic acid molecule comprises the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102.
147. An in vitro method of screening a sample from a subject for one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2), the method comprising
(a) isolating and purifying DNA, RNA, or protein (e.g., from a cell or population of cells) from a sample obtained from the subject; and
(b) detecting the presence or absence of one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA, RNA, or protein.
148. The method of any one of claims 105-147, wherein (a) comprises isolating and purifying DNA, or having DNA isolated and purified (e.g., from a cell or population of cells), from a sample obtained from the subject; and (b) comprises detecting, or having detected, the presence or absence of the one or more somatic gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) mutation (e.g., a somatic mutation set forth in Table 2) in the DNA.
149. The method of any one of claims 105-148, wherein the sample is a blood, tissue, or cell sample.
150. The method of any one of claims 105-149, wherein the sample is biopsy.
151. The method of any one of claims 105-150, wherein the sample comprises a cell (e.g., a T- cell) or a population of cells (e.g., a population of T-cells).
152. The method of any one of claims 105- 1 1 , wherein at least one of the one or more somatic mutation is a loss of function mutation, a gain of function mutation, a misscnsc mutation, synonymous, and/or non-synonymous.
153. The method of any one of claims 105-152, wherein the one or more somatic mutation is a loss of function mutation.
154. The method of any one of claims 105-153, wherein the one or more somatic mutation truncates the protein encoded by the gene.
155. The method of any one of claims 105-154, wherein the one or more somatic mutation is one or more somatic mutation set forth herein.
156. The method of any one of claims 105-155, wherein the one or more somatic mutation is one or more somatic mutation set forth in Table 2 herein.
157. A method of reducing or inhibiting expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell, the method comprising delivering into the cell the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell.
158. A method of reducing or inhibiting expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in a cell in a subject, the method comprising administering to the subject the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102, to thereby reduce or inhibit expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) in the cell in the subject.
159. The method of any one of claims 157-158, wherein the gene is BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6.
160. A method of treating, ameliorating, or preventing a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in a subject, the method comprising administering to the subject the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102, to thereby treat, ameliorate, or prevent the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease in the subject.
161. The method of claim 160, wherein the gene associated disease is a BTG1 associated disease, a CDKN1B associated disease, a DUSP2 associated disease, an IGLL5 associated disease, or a KLHL6 associated disease.
162. The method of any one of claims 160-161, wherein the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease associated disease is a proinflammatory disease (e.g., an autoimmune disease).
163. The method of any one of claims 160-162, wherein the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease associated disease is an autoimmune disease.
164. The method of any one of claims 160-163, wherein the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6) associated disease is treated, ameliorated, or prevented through the reduction or inhibition expression of the gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
165. A method of treating, ameliorate, or preventing a proinflammatory (e.g., autoimmune) disease in a subject, the method comprising administering to the subject the dsRNA agent of any one of claims 3-92, the conjugate of any one of claims 93-96, the vector of claim 97, the carrier of any one of claims 98-100, the cell of claim 101, or the pharmaceutical composition of claim 102, to thereby treat, ameliorate, or prevent the proinflammatory (e.g., autoimmune) disease in the subject.
166. The method of any claim 165, wherein the proinflammatory (e.g., autoimmune) disease is treated, ameliorated, or prevented through a reduction or inhibition of expression of a gene (e.g., BTG1, CDKN1B, DUSP2, IGLL5, or KLHL6).
167. The method of any one of claims 105-166, wherein the subject is a human.
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Citations (239)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US513030A (en) 1894-01-16 Machine for waxing or coating paper
US564562A (en) 1896-07-21 Joseph p
US3687808A (en) 1969-08-14 1972-08-29 Univ Leland Stanford Junior Synthetic polynucleotides
US4469863A (en) 1980-11-12 1984-09-04 Ts O Paul O P Nonionic nucleic acid alkyl and aryl phosphonates and processes for manufacture and use thereof
US4476301A (en) 1982-04-29 1984-10-09 Centre National De La Recherche Scientifique Oligonucleotides, a process for preparing the same and their application as mediators of the action of interferon
US4845205A (en) 1985-01-08 1989-07-04 Institut Pasteur 2,N6 -disubstituted and 2,N6 -trisubstituted adenosine-3'-phosphoramidites
US4981957A (en) 1984-07-19 1991-01-01 Centre National De La Recherche Scientifique Oligonucleotides with modified phosphate and modified carbohydrate moieties at the respective chain termini
US5023243A (en) 1981-10-23 1991-06-11 Molecular Biosystems, Inc. Oligonucleotide therapeutic agent and method of making same
US5034506A (en) 1985-03-15 1991-07-23 Anti-Gene Development Group Uncharged morpholino-based polymers having achiral intersubunit linkages
US5118800A (en) 1983-12-20 1992-06-02 California Institute Of Technology Oligonucleotides possessing a primary amino group in the terminal nucleotide
US5130302A (en) 1989-12-20 1992-07-14 Boron Bilogicals, Inc. Boronated nucleoside, nucleotide and oligonucleotide compounds, compositions and methods for using same
US5134066A (en) 1989-08-29 1992-07-28 Monsanto Company Improved probes using nucleosides containing 3-dezauracil analogs
US5166315A (en) 1989-12-20 1992-11-24 Anti-Gene Development Group Sequence-specific binding polymers for duplex nucleic acids
US5175273A (en) 1988-07-01 1992-12-29 Genentech, Inc. Nucleic acid intercalating agents
US5177195A (en) 1991-01-08 1993-01-05 Imperial Chemical Industries Plc Disazo dyes
US5185444A (en) 1985-03-15 1993-02-09 Anti-Gene Deveopment Group Uncharged morpolino-based polymers having phosphorous containing chiral intersubunit linkages
US5188897A (en) 1987-10-22 1993-02-23 Temple University Of The Commonwealth System Of Higher Education Encapsulated 2',5'-phosphorothioate oligoadenylates
US5214134A (en) 1990-09-12 1993-05-25 Sterling Winthrop Inc. Process of linking nucleosides with a siloxane bridge
US5216141A (en) 1988-06-06 1993-06-01 Benner Steven A Oligonucleotide analogs containing sulfur linkages
US5235033A (en) 1985-03-15 1993-08-10 Anti-Gene Development Group Alpha-morpholino ribonucleoside derivatives and polymers thereof
US5264564A (en) 1989-10-24 1993-11-23 Gilead Sciences Oligonucleotide analogs with novel linkages
US5264423A (en) 1987-03-25 1993-11-23 The United States Of America As Represented By The Department Of Health And Human Services Inhibitors for replication of retroviruses and for the expression of oncogene products
US5276019A (en) 1987-03-25 1994-01-04 The United States Of America As Represented By The Department Of Health And Human Services Inhibitors for replication of retroviruses and for the expression of oncogene products
US5278302A (en) 1988-05-26 1994-01-11 University Patents, Inc. Polynucleotide phosphorodithioates
US5319080A (en) 1991-10-17 1994-06-07 Ciba-Geigy Corporation Bicyclic nucleosides, oligonucleotides, process for their preparation and intermediates
US5321131A (en) 1990-03-08 1994-06-14 Hybridon, Inc. Site-specific functionalization of oligodeoxynucleotides for non-radioactive labelling
US5359044A (en) 1991-12-13 1994-10-25 Isis Pharmaceuticals Cyclobutyl oligonucleotide surrogates
US5367066A (en) 1984-10-16 1994-11-22 Chiron Corporation Oligonucleotides with selectably cleavable and/or abasic sites
US5399676A (en) 1989-10-23 1995-03-21 Gilead Sciences Oligonucleotides with inverted polarity
US5405938A (en) 1989-12-20 1995-04-11 Anti-Gene Development Group Sequence-specific binding polymers for duplex nucleic acids
US5405939A (en) 1987-10-22 1995-04-11 Temple University Of The Commonwealth System Of Higher Education 2',5'-phosphorothioate oligoadenylates and their covalent conjugates with polylysine
US5432272A (en) 1990-10-09 1995-07-11 Benner; Steven A. Method for incorporating into a DNA or RNA oligonucleotide using nucleotides bearing heterocyclic bases
US5434257A (en) 1992-06-01 1995-07-18 Gilead Sciences, Inc. Binding compentent oligomers containing unsaturated 3',5' and 2',5' linkages
US5446137A (en) 1993-12-09 1995-08-29 Syntex (U.S.A.) Inc. Oligonucleotides containing 4'-substituted nucleotides
US5455233A (en) 1989-11-30 1995-10-03 University Of North Carolina Oligoribonucleoside and oligodeoxyribonucleoside boranophosphates
US5457191A (en) 1990-01-11 1995-10-10 Isis Pharmaceuticals, Inc. 3-deazapurines
US5457187A (en) 1993-12-08 1995-10-10 Board Of Regents University Of Nebraska Oligonucleotides containing 5-fluorouracil
US5459255A (en) 1990-01-11 1995-10-17 Isis Pharmaceuticals, Inc. N-2 substituted purines
US5466786A (en) 1989-10-24 1995-11-14 Gilead Sciences 2'modified nucleoside and nucleotide compounds
US5466677A (en) 1993-03-06 1995-11-14 Ciba-Geigy Corporation Dinucleoside phosphinates and their pharmaceutical compositions
US5470967A (en) 1990-04-10 1995-11-28 The Dupont Merck Pharmaceutical Company Oligonucleotide analogs with sulfamate linkages
US5476925A (en) 1993-02-01 1995-12-19 Northwestern University Oligodeoxyribonucleotides including 3'-aminonucleoside-phosphoramidate linkages and terminal 3'-amino groups
US5484908A (en) 1991-11-26 1996-01-16 Gilead Sciences, Inc. Oligonucleotides containing 5-propynyl pyrimidines
US5489677A (en) 1990-07-27 1996-02-06 Isis Pharmaceuticals, Inc. Oligonucleoside linkages containing adjacent oxygen and nitrogen atoms
US5502177A (en) 1993-09-17 1996-03-26 Gilead Sciences, Inc. Pyrimidine derivatives for labeled binding partners
US5514785A (en) 1990-05-11 1996-05-07 Becton Dickinson And Company Solid supports for nucleic acid hybridization assays
US5519126A (en) 1988-03-25 1996-05-21 University Of Virginia Alumni Patents Foundation Oligonucleotide N-alkylphosphoramidates
US5519134A (en) 1994-01-11 1996-05-21 Isis Pharmaceuticals, Inc. Pyrrolidine-containing monomers and oligomers
US5525711A (en) 1994-05-18 1996-06-11 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Pteridine nucleotide analogs as fluorescent DNA probes
US5539082A (en) 1993-04-26 1996-07-23 Nielsen; Peter E. Peptide nucleic acids
US5541307A (en) 1990-07-27 1996-07-30 Isis Pharmaceuticals, Inc. Backbone modified oligonucleotide analogs and solid phase synthesis thereof
US5541316A (en) 1992-02-11 1996-07-30 Henkel Kommanditgesellschaft Auf Aktien Process for the production of polysaccharide-based polycarboxylates
US5550111A (en) 1984-07-11 1996-08-27 Temple University-Of The Commonwealth System Of Higher Education Dual action 2',5'-oligoadenylate antiviral derivatives and uses thereof
US5552540A (en) 1987-06-24 1996-09-03 Howard Florey Institute Of Experimental Physiology And Medicine Nucleoside derivatives
US5561225A (en) 1990-09-19 1996-10-01 Southern Research Institute Polynucleotide analogs containing sulfonate and sulfonamide internucleoside linkages
US5567811A (en) 1990-05-03 1996-10-22 Amersham International Plc Phosphoramidite derivatives, their preparation and the use thereof in the incorporation of reporter groups on synthetic oligonucleotides
US5571799A (en) 1991-08-12 1996-11-05 Basco, Ltd. (2'-5') oligoadenylate analogues useful as inhibitors of host-v5.-graft response
US5576427A (en) 1993-03-30 1996-11-19 Sterling Winthrop, Inc. Acyclic nucleoside analogs and oligonucleotide sequences containing them
US5587470A (en) 1990-01-11 1996-12-24 Isis Pharmaceuticals, Inc. 3-deazapurines
US5587361A (en) 1991-10-15 1996-12-24 Isis Pharmaceuticals, Inc. Oligonucleotides having phosphorothioate linkages of high chiral purity
US5591722A (en) 1989-09-15 1997-01-07 Southern Research Institute 2'-deoxy-4'-thioribonucleosides and their antiviral activity
US5594121A (en) 1991-11-07 1997-01-14 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified purines
US5596091A (en) 1994-03-18 1997-01-21 The Regents Of The University Of California Antisense oligonucleotides comprising 5-aminoalkyl pyrimidine nucleotides
US5596086A (en) 1990-09-20 1997-01-21 Gilead Sciences, Inc. Modified internucleoside linkages having one nitrogen and two carbon atoms
US5597909A (en) 1994-08-25 1997-01-28 Chiron Corporation Polynucleotide reagents containing modified deoxyribose moieties, and associated methods of synthesis and use
US5602240A (en) 1990-07-27 1997-02-11 Ciba Geigy Ag. Backbone modified oligonucleotide analogs
US5608046A (en) 1990-07-27 1997-03-04 Isis Pharmaceuticals, Inc. Conjugated 4'-desmethyl nucleoside analog compounds
US5610300A (en) 1992-07-01 1997-03-11 Ciba-Geigy Corporation Carbocyclic nucleosides containing bicyclic rings, oligonucleotides therefrom, process for their preparation, their use and intermediates
US5610289A (en) 1990-07-27 1997-03-11 Isis Pharmaceuticals, Inc. Backbone modified oligonucleotide analogues
US5614617A (en) 1990-07-27 1997-03-25 Isis Pharmaceuticals, Inc. Nuclease resistant, pyrimidine modified oligonucleotides that detect and modulate gene expression
US5618704A (en) 1990-07-27 1997-04-08 Isis Pharmacueticals, Inc. Backbone-modified oligonucleotide analogs and preparation thereof through radical coupling
US5623070A (en) 1990-07-27 1997-04-22 Isis Pharmaceuticals, Inc. Heteroatomic oligonucleoside linkages
US5625050A (en) 1994-03-31 1997-04-29 Amgen Inc. Modified oligonucleotides and intermediates useful in nucleic acid therapeutics
US5627053A (en) 1994-03-29 1997-05-06 Ribozyme Pharmaceuticals, Inc. 2'deoxy-2'-alkylnucleotide containing nucleic acid
US5633360A (en) 1992-04-14 1997-05-27 Gilead Sciences, Inc. Oligonucleotide analogs capable of passive cell membrane permeation
US5639873A (en) 1992-02-05 1997-06-17 Centre National De La Recherche Scientifique (Cnrs) Oligothionucleotides
US5645985A (en) 1991-11-26 1997-07-08 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified pyrimidines
US5646265A (en) 1990-01-11 1997-07-08 Isis Pharmceuticals, Inc. Process for the preparation of 2'-O-alkyl purine phosphoramidites
US5658873A (en) 1993-04-10 1997-08-19 Degussa Aktiengesellschaft Coated sodium percarbonate particles, a process for their production and detergent, cleaning and bleaching compositions containing them
US5663312A (en) 1993-03-31 1997-09-02 Sanofi Oligonucleotide dimers with amide linkages replacing phosphodiester linkages
US5670633A (en) 1990-01-11 1997-09-23 Isis Pharmaceuticals, Inc. Sugar modified oligonucleotides that detect and modulate gene expression
US5677437A (en) 1990-07-27 1997-10-14 Isis Pharmaceuticals, Inc. Heteroatomic oligonucleoside linkages
US5677439A (en) 1990-08-03 1997-10-14 Sanofi Oligonucleotide analogues containing phosphate diester linkage substitutes, compositions thereof, and precursor dinucleotide analogues
US5681941A (en) 1990-01-11 1997-10-28 Isis Pharmaceuticals, Inc. Substituted purines and oligonucleotide cross-linking
US5714331A (en) 1991-05-24 1998-02-03 Buchardt, Deceased; Ole Peptide nucleic acids having enhanced binding affinity, sequence specificity and solubility
US5719262A (en) 1993-11-22 1998-02-17 Buchardt, Deceased; Ole Peptide nucleic acids having amino acid side chains
US5739169A (en) 1996-05-31 1998-04-14 Procept, Incorporated Aromatic compounds for inhibiting immune response
US5801005A (en) 1993-03-17 1998-09-01 University Of Washington Immune reactivity to HER-2/neu protein for diagnosis of malignancies in which the HER-2/neu oncogene is associated
US5824311A (en) 1987-11-30 1998-10-20 Trustees Of The University Of Pennsylvania Treatment of tumors with monoclonal antibodies against oncogene antigens
US5830880A (en) 1994-08-26 1998-11-03 Hoechst Aktiengesellschaft Gene therapy of tumors with an endothelial cell-specific, cell cycle-dependent active compound
US5830653A (en) 1991-11-26 1998-11-03 Gilead Sciences, Inc. Methods of using oligomers containing modified pyrimidines
US5846945A (en) 1993-02-16 1998-12-08 Onyx Pharmaceuticals, Inc. Cytopathic viruses for therapy and prophylaxis of neoplasia
US5859221A (en) 1990-01-11 1999-01-12 Isis Pharmaceuticals, Inc. 2'-modified oligonucleotides
US5885613A (en) 1994-09-30 1999-03-23 The University Of British Columbia Bilayer stabilizing components and their use in forming programmable fusogenic liposomes
WO1999014226A2 (en) 1997-09-12 1999-03-25 Exiqon A/S Bi- and tri-cyclic nucleoside, nucleotide and oligonucleotide analogues
US5948903A (en) 1991-01-11 1999-09-07 Isis Pharmaceuticals, Inc. Synthesis of 3-deazapurines
US6005087A (en) 1995-06-06 1999-12-21 Isis Pharmaceuticals, Inc. 2'-modified oligonucleotides
US6015886A (en) 1993-05-24 2000-01-18 Chemgenes Corporation Oligonucleotide phosphate esters
US6028188A (en) 1993-11-16 2000-02-22 Genta Incorporated Synthetic oligomers having chirally pure phosphonate internucleosidyl linkages mixed with non-phosphonate internucleosidyl linkages
US6124445A (en) 1994-11-23 2000-09-26 Isis Pharmaceuticals, Inc. Phosphotriester oligonucleotides, amidities and method of preparation
US6147200A (en) 1999-08-19 2000-11-14 Isis Pharmaceuticals, Inc. 2'-O-acetamido modified monomers and oligomers
US6160109A (en) 1995-10-20 2000-12-12 Isis Pharmaceuticals, Inc. Preparation of phosphorothioate and boranophosphate oligomers
US6166197A (en) 1995-03-06 2000-12-26 Isis Pharmaceuticals, Inc. Oligomeric compounds having pyrimidine nucleotide (S) with 2'and 5 substitutions
US6169170B1 (en) 1994-03-18 2001-01-02 Lynx Therapeutics, Inc. Oligonucleotide N3′→N5′Phosphoramidate Duplexes
US6172209B1 (en) 1997-02-14 2001-01-09 Isis Pharmaceuticals Inc. Aminooxy-modified oligonucleotides and methods for making same
US6222025B1 (en) 1995-03-06 2001-04-24 Isis Pharmaceuticals, Inc. Process for the synthesis of 2′-O-substituted pyrimidines and oligomeric compounds therefrom
US6235887B1 (en) 1991-11-26 2001-05-22 Isis Pharmaceuticals, Inc. Enhanced triple-helix and double-helix formation directed by oligonucleotides containing modified pyrimidines
US6239265B1 (en) 1990-01-11 2001-05-29 Isis Pharmaceuticals, Inc. Oligonucleotides having chiral phosphorus linkages
US6268490B1 (en) 1997-03-07 2001-07-31 Takeshi Imanishi Bicyclonucleoside and oligonucleotide analogues
US6277603B1 (en) 1991-12-24 2001-08-21 Isis Pharmaceuticals, Inc. PNA-DNA-PNA chimeric macromolecules
US6287591B1 (en) 1997-05-14 2001-09-11 Inex Pharmaceuticals Corp. Charged therapeutic agents encapsulated in lipid particles containing four lipid components
US6326199B1 (en) 1991-12-24 2001-12-04 Isis Pharmaceuticals, Inc. Gapped 2′ modified oligonucleotides
US6346614B1 (en) 1992-07-23 2002-02-12 Hybridon, Inc. Hybrid oligonucleotide phosphorothioates
US6444423B1 (en) 1996-06-07 2002-09-03 Molecular Dynamics, Inc. Nucleosides comprising polydentate ligands
US6525191B1 (en) 1999-05-11 2003-02-25 Kanda S. Ramasamy Conformationally constrained L-nucleosides
US6528640B1 (en) 1997-11-05 2003-03-04 Ribozyme Pharmaceuticals, Incorporated Synthetic ribonucleic acids with RNAse activity
US6531590B1 (en) 1998-04-24 2003-03-11 Isis Pharmaceuticals, Inc. Processes for the synthesis of oligonucleotide compounds
US6534639B1 (en) 1999-07-07 2003-03-18 Isis Pharmaceuticals, Inc. Guanidinium functionalized oligonucleotides and method/synthesis
US20030077829A1 (en) 2001-04-30 2003-04-24 Protiva Biotherapeutics Inc.. Lipid-based formulations
US6608035B1 (en) 1994-10-25 2003-08-19 Hybridon, Inc. Method of down-regulating gene expression
US6617438B1 (en) 1997-11-05 2003-09-09 Sirna Therapeutics, Inc. Oligoribonucleotides with enzymatic activity
US6639062B2 (en) 1997-02-14 2003-10-28 Isis Pharmaceuticals, Inc. Aminooxy-modified nucleosidic compounds and oligomeric compounds prepared therefrom
US6670461B1 (en) 1997-09-12 2003-12-30 Exiqon A/S Oligonucleotide analogues
US6770748B2 (en) 1997-03-07 2004-08-03 Takeshi Imanishi Bicyclonucleoside and oligonucleotide analogue
WO2004106356A1 (en) 2003-05-27 2004-12-09 Syddansk Universitet Functionalized nucleotide derivatives
US6858715B2 (en) 1999-02-04 2005-02-22 Isis Pharmaceuticals, Inc. Process for the synthesis of oligomeric compounds
US6867294B1 (en) 1998-07-14 2005-03-15 Isis Pharmaceuticals, Inc. Gapped oligomers having site specific chiral phosphorothioate internucleoside linkages
US6878805B2 (en) 2002-08-16 2005-04-12 Isis Pharmaceuticals, Inc. Peptide-conjugated oligomeric compounds
WO2005071093A2 (en) 2004-01-23 2005-08-04 Istituto Di Ricerche Di Biologia Molecolare P Angeletti Spa Chimpanzee adenovirus vaccine carriers
US20050175682A1 (en) 2003-09-15 2005-08-11 Protiva Biotherapeutics, Inc. Polyethyleneglycol-modified lipid compounds and uses thereof
US20060008378A1 (en) 2004-04-30 2006-01-12 Kunihiro Imai Sterilization method
US6998484B2 (en) 2000-10-04 2006-02-14 Santaris Pharma A/S Synthesis of purine locked nucleic acid analogues
US7015315B1 (en) 1991-12-24 2006-03-21 Isis Pharmaceuticals, Inc. Gapped oligonucleotides
WO2006048215A1 (en) 2004-11-02 2006-05-11 Istituto Di Ricerche Di Biologia Molecolare P Angeletti Spa Adenoviral amplicon and producer cells for the production of replication-defective adenoviral vectors, methods of preparation and use thereof
US7045610B2 (en) 1998-04-03 2006-05-16 Epoch Biosciences, Inc. Modified oligonucleotides for mismatch discrimination
US7053207B2 (en) 1999-05-04 2006-05-30 Exiqon A/S L-ribo-LNA analogues
US7084125B2 (en) 1999-03-18 2006-08-01 Exiqon A/S Xylo-LNA analogues
US7273933B1 (en) 1998-02-26 2007-09-25 Isis Pharmaceuticals, Inc. Methods for synthesis of oligonucleotides
WO2007134181A2 (en) 2006-05-11 2007-11-22 Isis Pharmaceuticals, Inc. 5'-modified bicyclic nucleic acid analogs
US7321029B2 (en) 2000-01-21 2008-01-22 Geron Corporation 2′-arabino-fluorooligonucleotide N3′→P5′ phosphoramidates: their synthesis and use
US20080020058A1 (en) 2005-02-14 2008-01-24 Sirna Therapeutics, Inc. Lipid nanoparticle based compositions and methods for the delivery of biologically active molecules
US20080039618A1 (en) 2002-11-05 2008-02-14 Charles Allerson Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
US20080042973A1 (en) 2006-07-10 2008-02-21 Memsic, Inc. System for sensing yaw rate using a magnetic field sensor and portable electronic devices using the same
US7399845B2 (en) 2006-01-27 2008-07-15 Isis Pharmaceuticals, Inc. 6-modified bicyclic nucleic acid analogs
WO2008101157A1 (en) 2007-02-15 2008-08-21 Isis Pharmaceuticals, Inc. 5'-substituted-2'-f modified nucleosides and oligomeric compounds prepared therefrom
US7427672B2 (en) 2003-08-28 2008-09-23 Takeshi Imanishi Artificial nucleic acids of n-o bond crosslinkage type
US7495088B1 (en) 1989-12-04 2009-02-24 Enzo Life Sciences, Inc. Modified nucleotide compounds
US7569686B1 (en) 2006-01-27 2009-08-04 Isis Pharmaceuticals, Inc. Compounds and methods for synthesis of bicyclic nucleic acid analogs
US7572582B2 (en) 1997-09-12 2009-08-11 Exiqon A/S Oligonucleotide analogues
WO2009127060A1 (en) 2008-04-15 2009-10-22 Protiva Biotherapeutics, Inc. Novel lipid formulations for nucleic acid delivery
WO2009132131A1 (en) 2008-04-22 2009-10-29 Alnylam Pharmaceuticals, Inc. Amino lipid based improved lipid formulation
WO2009150222A2 (en) 2008-06-12 2009-12-17 Affitech A/S Improved protein expression system
US7666854B2 (en) 2006-05-11 2010-02-23 Isis Pharmaceuticals, Inc. Bis-modified bicyclic nucleic acid analogs
US20100062967A1 (en) 2004-12-27 2010-03-11 Silence Therapeutics Ag Coated lipid complexes and their use
US7696345B2 (en) 2002-11-05 2010-04-13 Isis Pharmaceuticals, Inc. Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
WO2010053572A2 (en) 2008-11-07 2010-05-14 Massachusetts Institute Of Technology Aminoalcohol lipidoids and uses thereof
WO2011005861A1 (en) 2009-07-07 2011-01-13 Alnylam Pharmaceuticals, Inc. Oligonucleotide end caps
US20110076335A1 (en) 2009-07-01 2011-03-31 Protiva Biotherapeutics, Inc. Novel lipid formulations for delivery of therapeutic agents to solid tumors
US20110117125A1 (en) 2008-01-02 2011-05-19 Tekmira Pharmaceuticals Corporation Compositions and methods for the delivery of nucleic acids
US20110256175A1 (en) 2008-10-09 2011-10-20 The University Of British Columbia Amino lipids and methods for the delivery of nucleic acids
US20110313020A1 (en) 2008-12-03 2011-12-22 Marina Biotech, Inc. UsiRNA Complexes
US20120011478A1 (en) 2005-02-24 2012-01-12 Texas Instruments Incorporated Merging sub-resolution assist features of a photolithographic mask
US20120027796A1 (en) 2008-11-10 2012-02-02 Alnylam Pharmaceuticals, Inc. Novel lipids and compositions for the delivery of therapeutics
US20120027803A1 (en) 2010-06-03 2012-02-02 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US20120149894A1 (en) 2009-08-20 2012-06-14 Mark Cameron Novel cationic lipids with various head groups for oligonucleotide delivery
US20120202871A1 (en) 2009-07-01 2012-08-09 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
US8278283B2 (en) 2007-07-05 2012-10-02 Isis Pharmaceuticals, Inc. 6-disubstituted or unsaturated bicyclic nucleic acid analogs
US8278425B2 (en) 2007-05-30 2012-10-02 Isis Pharmaceuticals, Inc. N-substituted-aminomethylene bridged bicyclic nucleic acid analogs
US8278426B2 (en) 2007-06-08 2012-10-02 Isis Pharmaceuticals, Inc. Carbocyclic bicyclic nucleic acid analogs
US20120287670A1 (en) 2010-04-09 2012-11-15 Sharp Kabushiki Kaisha Light source module and electronic apparatus provided with same
US8314227B2 (en) 2007-05-22 2012-11-20 Marina Biotech, Inc. Hydroxymethyl substituted RNA oligonucleotides and RNA complexes
WO2012162210A1 (en) 2011-05-26 2012-11-29 Merck Sharp & Dohme Corp. Ring constrained cationic lipids for oligonucleotide delivery
US20130011922A1 (en) 2007-03-02 2013-01-10 F/K/A Mdrna, Inc. Nucleic acid compounds for inhibiting gene expression and uses thereof
US20130022649A1 (en) 2009-12-01 2013-01-24 Protiva Biotherapeutics, Inc. Snalp formulations containing antioxidants
WO2013016058A1 (en) 2011-07-22 2013-01-31 Merck Sharp & Dohme Corp. Novel bis-nitrogen containing cationic lipids for oligonucleotide delivery
US20130053572A1 (en) 2010-01-22 2013-02-28 Steven L. Colletti Novel Cationic Lipids for Oligonucleotide Delivery
WO2013036868A1 (en) 2011-09-07 2013-03-14 Marina Biotech Inc. Synthesis and uses of nucleic acid compounds with conformationally restricted monomers
US20130090372A1 (en) 2010-06-04 2013-04-11 Brian W. Budzik Novel Low Molecular Weight Cationic Lipids for Oligonucleotide Delivery
US20130116307A1 (en) 2010-05-12 2013-05-09 Protiva Biotherapeutics Inc. Novel cyclic cationic lipids and methods of use
US20130123338A1 (en) 2010-05-12 2013-05-16 Protiva Biotherapeutics, Inc. Novel cationic lipids and methods of use thereof
WO2013075035A1 (en) 2011-11-18 2013-05-23 Alnylam Pharmaceuticals Rnai agents, compositions and methods of use thereof for treating transthyretin (ttr) associated diseases
US20130178541A1 (en) 2010-09-20 2013-07-11 Matthew G. Stanton Novel low molecular weight cationic lipids for oligonucleotide delivery
US20130189351A1 (en) 2010-08-31 2013-07-25 Novartis Ag Lipids suitable for liposomal delivery of protein coding rna
US20130190383A1 (en) 2010-04-26 2013-07-25 Marina Biotech, Inc. Nucleic acid compounds with conformationally restricted monomers and uses thereof
US20130195920A1 (en) 2011-12-07 2013-08-01 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US8501805B2 (en) 2008-09-24 2013-08-06 Isis Pharmaceuticals, Inc. Substituted alpha-L-bicyclic nucleosides
US20130203836A1 (en) 2010-04-01 2013-08-08 Isis Pharmaceuticals, Inc. 2' and 5' modified monomers and oligonucleotides
WO2013116126A1 (en) 2012-02-01 2013-08-08 Merck Sharp & Dohme Corp. Novel low molecular weight, biodegradable cationic lipids for oligonucleotide delivery
US8530640B2 (en) 2008-02-07 2013-09-10 Isis Pharmaceuticals, Inc. Bicyclic cyclohexitol nucleic acid analogs
US8546556B2 (en) 2007-11-21 2013-10-01 Isis Pharmaceuticals, Inc Carbocyclic alpha-L-bicyclic nucleic acid analogs
US20130274504A1 (en) 2010-10-21 2013-10-17 Steven L. Colletti Novel Low Molecular Weight Cationic Lipids For Oligonucleotide Delivery
US20130274523A1 (en) 2010-09-30 2013-10-17 John A. Bawiec, III Low molecular weight cationic lipids for oligonucleotide delivery
US20130303587A1 (en) 2010-06-30 2013-11-14 Protiva Biotherapeutics, Inc. Non-liposomal systems for nucleic acid delivery
US20130323269A1 (en) 2010-07-30 2013-12-05 Muthiah Manoharan Methods and compositions for delivery of active agents
US20130338210A1 (en) 2009-12-07 2013-12-19 Alnylam Pharmaceuticals, Inc. Compositions for nucleic acid delivery
US20140039032A1 (en) 2011-12-12 2014-02-06 Kyowa Hakko Kirin Co., Ltd. Lipid nano particles comprising cationic lipid for drug delivery system
USRE44779E1 (en) 1997-03-07 2014-02-25 Santaris Pharma A/S Bicyclonucleoside and oligonucleotide analogues
WO2014062811A2 (en) 2012-10-16 2014-04-24 New York University METHOD OF TREATING CANCER WITH MODULATORS OF SCFSkp2
US20140200257A1 (en) 2011-01-11 2014-07-17 Alnylam Pharmaceuticals, Inc. Pegylated lipids and their use for drug delivery
WO2014136086A1 (en) 2013-03-08 2014-09-12 Novartis Ag Lipids and lipid compositions for the delivery of active agents
US20140308304A1 (en) 2011-12-07 2014-10-16 Alnylam Pharmaceuticals, Inc. Lipids for the delivery of active agents
US20150005363A1 (en) 2011-12-07 2015-01-01 Alnylam Pharmaceuticals, Inc. Branched Alkyl And Cycloalkyl Terminated Biodegradable Lipids For The Delivery Of Active Agents
WO2015016718A1 (en) 2013-08-02 2015-02-05 Bionovion Holding B.V. Combining cd27 agonists and immune checkpoint inhibition for immune stimulation
US20150057373A1 (en) 2012-03-27 2015-02-26 Sirna Therapeutics, Inc DIETHER BASED BIODEGRADABLE CATIONIC LIPIDS FOR siRNA DELIVERY
US20150064242A1 (en) 2012-02-24 2015-03-05 Protiva Biotherapeutics, Inc. Trialkyl cationic lipids and methods of use thereof
US9012421B2 (en) 2009-08-06 2015-04-21 Isis Pharmaceuticals, Inc. Bicyclic cyclohexose nucleic acid analogs
WO2015074085A1 (en) 2013-11-18 2015-05-21 Arcturus Therapeutics, Inc. Ionizable cationic lipid for rna delivery
US20150140070A1 (en) 2013-10-22 2015-05-21 Shire Human Genetic Therapies, Inc. Lipid formulations for delivery of messenger rna
WO2015095340A1 (en) 2013-12-19 2015-06-25 Novartis Ag Lipids and lipid compositions for the delivery of active agents
WO2015106128A2 (en) 2014-01-09 2015-07-16 Alnylam Pharmaceuticals, Inc. MODIFIED RNAi AGENTS
US20150203446A1 (en) 2011-09-27 2015-07-23 Takeda Pharmaceutical Company Limited Di-aliphatic substituted pegylated lipids
US20150239926A1 (en) 2013-11-18 2015-08-27 Arcturus Therapeutics, Inc. Asymmetric ionizable cationic lipid for rna delivery
WO2015199952A1 (en) 2014-06-25 2015-12-30 Acuitas Therapeutics Inc. Novel lipids and lipid nanoparticle formulations for delivery of nucleic acids
US20160151284A1 (en) 2013-07-23 2016-06-02 Protiva Biotherapeutics, Inc. Compositions and methods for delivering messenger rna
US20160317458A1 (en) 2013-12-19 2016-11-03 Luis Brito Lipids and Lipid Compositions for the Delivery of Active Agents
US20160376224A1 (en) 2015-06-29 2016-12-29 Acuitas Therapeutics, Inc. Lipids and lipid nanoparticle formulations for delivery of nucleic acids
US20170119904A1 (en) 2015-10-28 2017-05-04 Acuitas Therapeutics, Inc. Novel lipids and lipid nanoparticle formulations for delivery of nucleic acids
WO2017099823A1 (en) 2015-12-10 2017-06-15 Modernatx, Inc. Compositions and methods for delivery of therapeutic agents
WO2017117528A1 (en) 2015-12-30 2017-07-06 Acuitas Therapeutics, Inc. Lipids and lipid nanoparticle formulations for delivery of nucleic acids
US9708628B2 (en) 2011-11-18 2017-07-18 Nof Corporation Cationic lipid having improved intracellular kinetics
US20170210967A1 (en) 2010-12-06 2017-07-27 Schlumberger Technology Corporation Compositions and methods for well completions
WO2017173054A1 (en) 2016-03-30 2017-10-05 Intellia Therapeutics, Inc. Lipid nanoparticle formulations for crispr/cas components
US9867888B2 (en) 2015-09-17 2018-01-16 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
WO2018081480A1 (en) 2016-10-26 2018-05-03 Acuitas Therapeutics, Inc. Lipid nanoparticle formulations
US10086013B2 (en) 2011-10-27 2018-10-02 Massachusetts Institute Of Technology Amino acid-, peptide- and polypeptide-lipids, isomers, compositions, and uses thereof
WO2019051289A1 (en) 2017-09-08 2019-03-14 Generation Bio Co. Lipid nanoparticle formulations of non-viral, capsid-free dna vectors
WO2019067992A1 (en) 2017-09-29 2019-04-04 Intellia Therapeutics, Inc. Formulations
US20190240349A1 (en) 2015-06-19 2019-08-08 Massachusetts Institute Of Technology Alkenyl substituted 2,5-piperazinediones, compositions, and uses thereof
WO2019157531A1 (en) 2018-02-12 2019-08-15 Ionis Pharmaceuticals, Inc. Modified compounds and uses thereof
WO2019217941A1 (en) 2018-05-11 2019-11-14 Beam Therapeutics Inc. Methods of suppressing pathogenic mutations using programmable base editor systems
WO2020081938A1 (en) 2018-10-18 2020-04-23 Acuitas Therapeutics, Inc. Lipids for lipid nanoparticle delivery of active agents
WO2020106946A1 (en) 2018-11-21 2020-05-28 Translate Bio, Inc. TREATMENT OF CYSTIC FIBROSIS BY DELIVERY OF NEBULIZED mRNA ENCODING CFTR
WO2020219876A1 (en) 2019-04-25 2020-10-29 Intellia Therapeutics, Inc. Ionizable amine lipids and lipid nanoparticles
WO2021113777A2 (en) 2019-12-04 2021-06-10 Orna Therapeutics, Inc. Circular rna compositions and methods
WO2021257782A1 (en) 2020-06-18 2021-12-23 Alnylam Pharmaceuticals, Inc. XANTHINE DEHYDROGENASE (XDH) iRNA COMPOSITIONS AND METHODS OF USE THEREOF
US11446344B1 (en) 2018-12-12 2022-09-20 Flagship Pioneering Innovations V, Inc. Anellovirus compositions and methods of use
WO2022246251A2 (en) 2021-05-21 2022-11-24 Ionis Pharmaceuticals, Inc. Compounds for modulating unc13a expression
WO2022271573A1 (en) 2021-06-21 2022-12-29 Arrowhead Pharmaceuticals, Inc. Rnai agents for inhibiting expression of xanthine dehydrogenase (xdh), pharmaceutical compositions thereof, and methods of use
WO2024049971A2 (en) 2022-09-02 2024-03-07 Baylor College Of Medicine Modulation of b-cell translocation gene 1 (btg1) for use in adoptive cell therapy

Patent Citations (284)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US564562A (en) 1896-07-21 Joseph p
US513030A (en) 1894-01-16 Machine for waxing or coating paper
US3687808A (en) 1969-08-14 1972-08-29 Univ Leland Stanford Junior Synthetic polynucleotides
US4469863A (en) 1980-11-12 1984-09-04 Ts O Paul O P Nonionic nucleic acid alkyl and aryl phosphonates and processes for manufacture and use thereof
US5023243A (en) 1981-10-23 1991-06-11 Molecular Biosystems, Inc. Oligonucleotide therapeutic agent and method of making same
US4476301A (en) 1982-04-29 1984-10-09 Centre National De La Recherche Scientifique Oligonucleotides, a process for preparing the same and their application as mediators of the action of interferon
US5118800A (en) 1983-12-20 1992-06-02 California Institute Of Technology Oligonucleotides possessing a primary amino group in the terminal nucleotide
US5550111A (en) 1984-07-11 1996-08-27 Temple University-Of The Commonwealth System Of Higher Education Dual action 2',5'-oligoadenylate antiviral derivatives and uses thereof
US4981957A (en) 1984-07-19 1991-01-01 Centre National De La Recherche Scientifique Oligonucleotides with modified phosphate and modified carbohydrate moieties at the respective chain termini
US5367066A (en) 1984-10-16 1994-11-22 Chiron Corporation Oligonucleotides with selectably cleavable and/or abasic sites
US4845205A (en) 1985-01-08 1989-07-04 Institut Pasteur 2,N6 -disubstituted and 2,N6 -trisubstituted adenosine-3'-phosphoramidites
US5034506A (en) 1985-03-15 1991-07-23 Anti-Gene Development Group Uncharged morpholino-based polymers having achiral intersubunit linkages
US5185444A (en) 1985-03-15 1993-02-09 Anti-Gene Deveopment Group Uncharged morpolino-based polymers having phosphorous containing chiral intersubunit linkages
US5235033A (en) 1985-03-15 1993-08-10 Anti-Gene Development Group Alpha-morpholino ribonucleoside derivatives and polymers thereof
US5276019A (en) 1987-03-25 1994-01-04 The United States Of America As Represented By The Department Of Health And Human Services Inhibitors for replication of retroviruses and for the expression of oncogene products
US5264423A (en) 1987-03-25 1993-11-23 The United States Of America As Represented By The Department Of Health And Human Services Inhibitors for replication of retroviruses and for the expression of oncogene products
US5286717A (en) 1987-03-25 1994-02-15 The United States Of America As Represented By The Department Of Health And Human Services Inhibitors for replication of retroviruses and for the expression of oncogene products
US5552540A (en) 1987-06-24 1996-09-03 Howard Florey Institute Of Experimental Physiology And Medicine Nucleoside derivatives
US5405939A (en) 1987-10-22 1995-04-11 Temple University Of The Commonwealth System Of Higher Education 2',5'-phosphorothioate oligoadenylates and their covalent conjugates with polylysine
US5188897A (en) 1987-10-22 1993-02-23 Temple University Of The Commonwealth System Of Higher Education Encapsulated 2',5'-phosphorothioate oligoadenylates
US5824311A (en) 1987-11-30 1998-10-20 Trustees Of The University Of Pennsylvania Treatment of tumors with monoclonal antibodies against oncogene antigens
US5519126A (en) 1988-03-25 1996-05-21 University Of Virginia Alumni Patents Foundation Oligonucleotide N-alkylphosphoramidates
US5453496A (en) 1988-05-26 1995-09-26 University Patents, Inc. Polynucleotide phosphorodithioate
US5278302A (en) 1988-05-26 1994-01-11 University Patents, Inc. Polynucleotide phosphorodithioates
US5216141A (en) 1988-06-06 1993-06-01 Benner Steven A Oligonucleotide analogs containing sulfur linkages
US5175273A (en) 1988-07-01 1992-12-29 Genentech, Inc. Nucleic acid intercalating agents
US5134066A (en) 1989-08-29 1992-07-28 Monsanto Company Improved probes using nucleosides containing 3-dezauracil analogs
US5591722A (en) 1989-09-15 1997-01-07 Southern Research Institute 2'-deoxy-4'-thioribonucleosides and their antiviral activity
US5399676A (en) 1989-10-23 1995-03-21 Gilead Sciences Oligonucleotides with inverted polarity
US5264564A (en) 1989-10-24 1993-11-23 Gilead Sciences Oligonucleotide analogs with novel linkages
US5466786B1 (en) 1989-10-24 1998-04-07 Gilead Sciences 2' Modified nucleoside and nucleotide compounds
US5466786A (en) 1989-10-24 1995-11-14 Gilead Sciences 2'modified nucleoside and nucleotide compounds
US5455233A (en) 1989-11-30 1995-10-03 University Of North Carolina Oligoribonucleoside and oligodeoxyribonucleoside boranophosphates
US7495088B1 (en) 1989-12-04 2009-02-24 Enzo Life Sciences, Inc. Modified nucleotide compounds
US5130302A (en) 1989-12-20 1992-07-14 Boron Bilogicals, Inc. Boronated nucleoside, nucleotide and oligonucleotide compounds, compositions and methods for using same
US5405938A (en) 1989-12-20 1995-04-11 Anti-Gene Development Group Sequence-specific binding polymers for duplex nucleic acids
US5166315A (en) 1989-12-20 1992-11-24 Anti-Gene Development Group Sequence-specific binding polymers for duplex nucleic acids
US5750692A (en) 1990-01-11 1998-05-12 Isis Pharmaceuticals, Inc. Synthesis of 3-deazapurines
US5587470A (en) 1990-01-11 1996-12-24 Isis Pharmaceuticals, Inc. 3-deazapurines
US6166199A (en) 1990-01-11 2000-12-26 Isis Pharmaceuticals, Inc N-2 substituted purines
US5459255A (en) 1990-01-11 1995-10-17 Isis Pharmaceuticals, Inc. N-2 substituted purines
US5587469A (en) 1990-01-11 1996-12-24 Isis Pharmaceuticals, Inc. Oligonucleotides containing N-2 substituted purines
US6239265B1 (en) 1990-01-11 2001-05-29 Isis Pharmaceuticals, Inc. Oligonucleotides having chiral phosphorus linkages
US5646265A (en) 1990-01-11 1997-07-08 Isis Pharmceuticals, Inc. Process for the preparation of 2'-O-alkyl purine phosphoramidites
US5670633A (en) 1990-01-11 1997-09-23 Isis Pharmaceuticals, Inc. Sugar modified oligonucleotides that detect and modulate gene expression
US5859221A (en) 1990-01-11 1999-01-12 Isis Pharmaceuticals, Inc. 2'-modified oligonucleotides
US6531584B1 (en) 1990-01-11 2003-03-11 Isis Pharmaceuticals, Inc. 2'modified oligonucleotides
US5681941A (en) 1990-01-11 1997-10-28 Isis Pharmaceuticals, Inc. Substituted purines and oligonucleotide cross-linking
US5808027A (en) 1990-01-11 1998-09-15 Isis Pharmaceuticals, Inc. N-2 substituted purines in oligonucleotides
US5457191A (en) 1990-01-11 1995-10-10 Isis Pharmaceuticals, Inc. 3-deazapurines
US5321131A (en) 1990-03-08 1994-06-14 Hybridon, Inc. Site-specific functionalization of oligodeoxynucleotides for non-radioactive labelling
US5536821A (en) 1990-03-08 1996-07-16 Worcester Foundation For Biomedical Research Aminoalkylphosphorothioamidate oligonucleotide deratives
US5563253A (en) 1990-03-08 1996-10-08 Worcester Foundation For Biomedical Research Linear aminoalkylphosphoramidate oligonucleotide derivatives
US5470967A (en) 1990-04-10 1995-11-28 The Dupont Merck Pharmaceutical Company Oligonucleotide analogs with sulfamate linkages
US5567811A (en) 1990-05-03 1996-10-22 Amersham International Plc Phosphoramidite derivatives, their preparation and the use thereof in the incorporation of reporter groups on synthetic oligonucleotides
US5514785A (en) 1990-05-11 1996-05-07 Becton Dickinson And Company Solid supports for nucleic acid hybridization assays
US5618704A (en) 1990-07-27 1997-04-08 Isis Pharmacueticals, Inc. Backbone-modified oligonucleotide analogs and preparation thereof through radical coupling
US5602240A (en) 1990-07-27 1997-02-11 Ciba Geigy Ag. Backbone modified oligonucleotide analogs
US5677437A (en) 1990-07-27 1997-10-14 Isis Pharmaceuticals, Inc. Heteroatomic oligonucleoside linkages
US5608046A (en) 1990-07-27 1997-03-04 Isis Pharmaceuticals, Inc. Conjugated 4'-desmethyl nucleoside analog compounds
US5541307A (en) 1990-07-27 1996-07-30 Isis Pharmaceuticals, Inc. Backbone modified oligonucleotide analogs and solid phase synthesis thereof
US5610289A (en) 1990-07-27 1997-03-11 Isis Pharmaceuticals, Inc. Backbone modified oligonucleotide analogues
US5614617A (en) 1990-07-27 1997-03-25 Isis Pharmaceuticals, Inc. Nuclease resistant, pyrimidine modified oligonucleotides that detect and modulate gene expression
US5489677A (en) 1990-07-27 1996-02-06 Isis Pharmaceuticals, Inc. Oligonucleoside linkages containing adjacent oxygen and nitrogen atoms
US5623070A (en) 1990-07-27 1997-04-22 Isis Pharmaceuticals, Inc. Heteroatomic oligonucleoside linkages
US5677439A (en) 1990-08-03 1997-10-14 Sanofi Oligonucleotide analogues containing phosphate diester linkage substitutes, compositions thereof, and precursor dinucleotide analogues
US5214134A (en) 1990-09-12 1993-05-25 Sterling Winthrop Inc. Process of linking nucleosides with a siloxane bridge
US5561225A (en) 1990-09-19 1996-10-01 Southern Research Institute Polynucleotide analogs containing sulfonate and sulfonamide internucleoside linkages
US5596086A (en) 1990-09-20 1997-01-21 Gilead Sciences, Inc. Modified internucleoside linkages having one nitrogen and two carbon atoms
US5432272A (en) 1990-10-09 1995-07-11 Benner; Steven A. Method for incorporating into a DNA or RNA oligonucleotide using nucleotides bearing heterocyclic bases
US5177195A (en) 1991-01-08 1993-01-05 Imperial Chemical Industries Plc Disazo dyes
US5948903A (en) 1991-01-11 1999-09-07 Isis Pharmaceuticals, Inc. Synthesis of 3-deazapurines
US5714331A (en) 1991-05-24 1998-02-03 Buchardt, Deceased; Ole Peptide nucleic acids having enhanced binding affinity, sequence specificity and solubility
US5571799A (en) 1991-08-12 1996-11-05 Basco, Ltd. (2'-5') oligoadenylate analogues useful as inhibitors of host-v5.-graft response
US5587361A (en) 1991-10-15 1996-12-24 Isis Pharmaceuticals, Inc. Oligonucleotides having phosphorothioate linkages of high chiral purity
US5393878A (en) 1991-10-17 1995-02-28 Ciba-Geigy Corporation Bicyclic nucleosides, oligonucleotides, process for their preparation and intermediates
US5319080A (en) 1991-10-17 1994-06-07 Ciba-Geigy Corporation Bicyclic nucleosides, oligonucleotides, process for their preparation and intermediates
US5594121A (en) 1991-11-07 1997-01-14 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified purines
US5830653A (en) 1991-11-26 1998-11-03 Gilead Sciences, Inc. Methods of using oligomers containing modified pyrimidines
US5645985A (en) 1991-11-26 1997-07-08 Gilead Sciences, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified pyrimidines
US5484908A (en) 1991-11-26 1996-01-16 Gilead Sciences, Inc. Oligonucleotides containing 5-propynyl pyrimidines
US6235887B1 (en) 1991-11-26 2001-05-22 Isis Pharmaceuticals, Inc. Enhanced triple-helix and double-helix formation directed by oligonucleotides containing modified pyrimidines
US6380368B1 (en) 1991-11-26 2002-04-30 Isis Pharmaceuticals, Inc. Enhanced triple-helix and double-helix formation with oligomers containing modified pyrimidines
US5359044A (en) 1991-12-13 1994-10-25 Isis Pharmaceuticals Cyclobutyl oligonucleotide surrogates
US6277603B1 (en) 1991-12-24 2001-08-21 Isis Pharmaceuticals, Inc. PNA-DNA-PNA chimeric macromolecules
US6326199B1 (en) 1991-12-24 2001-12-04 Isis Pharmaceuticals, Inc. Gapped 2′ modified oligonucleotides
US7015315B1 (en) 1991-12-24 2006-03-21 Isis Pharmaceuticals, Inc. Gapped oligonucleotides
US5639873A (en) 1992-02-05 1997-06-17 Centre National De La Recherche Scientifique (Cnrs) Oligothionucleotides
US5541316A (en) 1992-02-11 1996-07-30 Henkel Kommanditgesellschaft Auf Aktien Process for the production of polysaccharide-based polycarboxylates
US5633360A (en) 1992-04-14 1997-05-27 Gilead Sciences, Inc. Oligonucleotide analogs capable of passive cell membrane permeation
US5434257A (en) 1992-06-01 1995-07-18 Gilead Sciences, Inc. Binding compentent oligomers containing unsaturated 3',5' and 2',5' linkages
US5610300A (en) 1992-07-01 1997-03-11 Ciba-Geigy Corporation Carbocyclic nucleosides containing bicyclic rings, oligonucleotides therefrom, process for their preparation, their use and intermediates
US5700920A (en) 1992-07-01 1997-12-23 Novartis Corporation Carbocyclic nucleosides containing bicyclic rings, oligonucleotides therefrom, process for their preparation, their use and intermediates
US6346614B1 (en) 1992-07-23 2002-02-12 Hybridon, Inc. Hybrid oligonucleotide phosphorothioates
US6683167B2 (en) 1992-07-23 2004-01-27 University Of Massachusetts Worcester Hybrid oligonucleotide phosphorothioates
US5476925A (en) 1993-02-01 1995-12-19 Northwestern University Oligodeoxyribonucleotides including 3'-aminonucleoside-phosphoramidate linkages and terminal 3'-amino groups
US5846945A (en) 1993-02-16 1998-12-08 Onyx Pharmaceuticals, Inc. Cytopathic viruses for therapy and prophylaxis of neoplasia
US5466677A (en) 1993-03-06 1995-11-14 Ciba-Geigy Corporation Dinucleoside phosphinates and their pharmaceutical compositions
US5801005A (en) 1993-03-17 1998-09-01 University Of Washington Immune reactivity to HER-2/neu protein for diagnosis of malignancies in which the HER-2/neu oncogene is associated
US5576427A (en) 1993-03-30 1996-11-19 Sterling Winthrop, Inc. Acyclic nucleoside analogs and oligonucleotide sequences containing them
US5663312A (en) 1993-03-31 1997-09-02 Sanofi Oligonucleotide dimers with amide linkages replacing phosphodiester linkages
US5658873A (en) 1993-04-10 1997-08-19 Degussa Aktiengesellschaft Coated sodium percarbonate particles, a process for their production and detergent, cleaning and bleaching compositions containing them
US5539082A (en) 1993-04-26 1996-07-23 Nielsen; Peter E. Peptide nucleic acids
US6015886A (en) 1993-05-24 2000-01-18 Chemgenes Corporation Oligonucleotide phosphate esters
US5502177A (en) 1993-09-17 1996-03-26 Gilead Sciences, Inc. Pyrimidine derivatives for labeled binding partners
US5763588A (en) 1993-09-17 1998-06-09 Gilead Sciences, Inc. Pyrimidine derivatives for labeled binding partners
US6005096A (en) 1993-09-17 1999-12-21 Gilead Sciences, Inc. Pyrimidine derivatives
US6028188A (en) 1993-11-16 2000-02-22 Genta Incorporated Synthetic oligomers having chirally pure phosphonate internucleosidyl linkages mixed with non-phosphonate internucleosidyl linkages
US5719262A (en) 1993-11-22 1998-02-17 Buchardt, Deceased; Ole Peptide nucleic acids having amino acid side chains
US5457187A (en) 1993-12-08 1995-10-10 Board Of Regents University Of Nebraska Oligonucleotides containing 5-fluorouracil
US5446137A (en) 1993-12-09 1995-08-29 Syntex (U.S.A.) Inc. Oligonucleotides containing 4'-substituted nucleotides
US5446137B1 (en) 1993-12-09 1998-10-06 Behringwerke Ag Oligonucleotides containing 4'-substituted nucleotides
US5519134A (en) 1994-01-11 1996-05-21 Isis Pharmaceuticals, Inc. Pyrrolidine-containing monomers and oligomers
US5811534A (en) 1994-02-01 1998-09-22 Isis Pharmaceuticals, Inc. Substituted purines and oligonucleotide cross-linking
US5596091A (en) 1994-03-18 1997-01-21 The Regents Of The University Of California Antisense oligonucleotides comprising 5-aminoalkyl pyrimidine nucleotides
US6169170B1 (en) 1994-03-18 2001-01-02 Lynx Therapeutics, Inc. Oligonucleotide N3′→N5′Phosphoramidate Duplexes
US5627053A (en) 1994-03-29 1997-05-06 Ribozyme Pharmaceuticals, Inc. 2'deoxy-2'-alkylnucleotide containing nucleic acid
US5625050A (en) 1994-03-31 1997-04-29 Amgen Inc. Modified oligonucleotides and intermediates useful in nucleic acid therapeutics
US5525711A (en) 1994-05-18 1996-06-11 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Pteridine nucleotide analogs as fluorescent DNA probes
US5597909A (en) 1994-08-25 1997-01-28 Chiron Corporation Polynucleotide reagents containing modified deoxyribose moieties, and associated methods of synthesis and use
US5830880A (en) 1994-08-26 1998-11-03 Hoechst Aktiengesellschaft Gene therapy of tumors with an endothelial cell-specific, cell cycle-dependent active compound
US5885613A (en) 1994-09-30 1999-03-23 The University Of British Columbia Bilayer stabilizing components and their use in forming programmable fusogenic liposomes
US6608035B1 (en) 1994-10-25 2003-08-19 Hybridon, Inc. Method of down-regulating gene expression
US6124445A (en) 1994-11-23 2000-09-26 Isis Pharmaceuticals, Inc. Phosphotriester oligonucleotides, amidities and method of preparation
US6222025B1 (en) 1995-03-06 2001-04-24 Isis Pharmaceuticals, Inc. Process for the synthesis of 2′-O-substituted pyrimidines and oligomeric compounds therefrom
US6166197A (en) 1995-03-06 2000-12-26 Isis Pharmaceuticals, Inc. Oligomeric compounds having pyrimidine nucleotide (S) with 2'and 5 substitutions
US6005087A (en) 1995-06-06 1999-12-21 Isis Pharmaceuticals, Inc. 2'-modified oligonucleotides
US6160109A (en) 1995-10-20 2000-12-12 Isis Pharmaceuticals, Inc. Preparation of phosphorothioate and boranophosphate oligomers
US5739169A (en) 1996-05-31 1998-04-14 Procept, Incorporated Aromatic compounds for inhibiting immune response
US6444423B1 (en) 1996-06-07 2002-09-03 Molecular Dynamics, Inc. Nucleosides comprising polydentate ligands
US6172209B1 (en) 1997-02-14 2001-01-09 Isis Pharmaceuticals Inc. Aminooxy-modified oligonucleotides and methods for making same
US6639062B2 (en) 1997-02-14 2003-10-28 Isis Pharmaceuticals, Inc. Aminooxy-modified nucleosidic compounds and oligomeric compounds prepared therefrom
US6268490B1 (en) 1997-03-07 2001-07-31 Takeshi Imanishi Bicyclonucleoside and oligonucleotide analogues
US6770748B2 (en) 1997-03-07 2004-08-03 Takeshi Imanishi Bicyclonucleoside and oligonucleotide analogue
USRE44779E1 (en) 1997-03-07 2014-02-25 Santaris Pharma A/S Bicyclonucleoside and oligonucleotide analogues
US6287591B1 (en) 1997-05-14 2001-09-11 Inex Pharmaceuticals Corp. Charged therapeutic agents encapsulated in lipid particles containing four lipid components
US6794499B2 (en) 1997-09-12 2004-09-21 Exiqon A/S Oligonucleotide analogues
WO1999014226A2 (en) 1997-09-12 1999-03-25 Exiqon A/S Bi- and tri-cyclic nucleoside, nucleotide and oligonucleotide analogues
US8153365B2 (en) 1997-09-12 2012-04-10 Exiqon A/S Oligonucleotide analogues
US6670461B1 (en) 1997-09-12 2003-12-30 Exiqon A/S Oligonucleotide analogues
US8080644B2 (en) 1997-09-12 2011-12-20 Exiqon A/S Oligonucleotide analogues
US7034133B2 (en) 1997-09-12 2006-04-25 Exiqon A/S Oligonucleotide analogues
US8034909B2 (en) 1997-09-12 2011-10-11 Exiqon A/S Oligonucleotide analogues
US7572582B2 (en) 1997-09-12 2009-08-11 Exiqon A/S Oligonucleotide analogues
US6617438B1 (en) 1997-11-05 2003-09-09 Sirna Therapeutics, Inc. Oligoribonucleotides with enzymatic activity
US6528640B1 (en) 1997-11-05 2003-03-04 Ribozyme Pharmaceuticals, Incorporated Synthetic ribonucleic acids with RNAse activity
US7273933B1 (en) 1998-02-26 2007-09-25 Isis Pharmaceuticals, Inc. Methods for synthesis of oligonucleotides
US7045610B2 (en) 1998-04-03 2006-05-16 Epoch Biosciences, Inc. Modified oligonucleotides for mismatch discrimination
US6531590B1 (en) 1998-04-24 2003-03-11 Isis Pharmaceuticals, Inc. Processes for the synthesis of oligonucleotide compounds
US6867294B1 (en) 1998-07-14 2005-03-15 Isis Pharmaceuticals, Inc. Gapped oligomers having site specific chiral phosphorothioate internucleoside linkages
USRE39464E1 (en) 1998-07-14 2007-01-09 Isis Pharmaceuticals Inc. Oligonucleolotides having site specific chiral phosphorothioate internucleoside linkages
US6858715B2 (en) 1999-02-04 2005-02-22 Isis Pharmaceuticals, Inc. Process for the synthesis of oligomeric compounds
US7041816B2 (en) 1999-02-04 2006-05-09 Isis Pharmaceuticals, Inc. Process for the synthesis of oligomeric compounds
US7084125B2 (en) 1999-03-18 2006-08-01 Exiqon A/S Xylo-LNA analogues
US7053207B2 (en) 1999-05-04 2006-05-30 Exiqon A/S L-ribo-LNA analogues
US6525191B1 (en) 1999-05-11 2003-02-25 Kanda S. Ramasamy Conformationally constrained L-nucleosides
US6534639B1 (en) 1999-07-07 2003-03-18 Isis Pharmaceuticals, Inc. Guanidinium functionalized oligonucleotides and method/synthesis
US6147200A (en) 1999-08-19 2000-11-14 Isis Pharmaceuticals, Inc. 2'-O-acetamido modified monomers and oligomers
US7321029B2 (en) 2000-01-21 2008-01-22 Geron Corporation 2′-arabino-fluorooligonucleotide N3′→P5′ phosphoramidates: their synthesis and use
US6998484B2 (en) 2000-10-04 2006-02-14 Santaris Pharma A/S Synthesis of purine locked nucleic acid analogues
US20030077829A1 (en) 2001-04-30 2003-04-24 Protiva Biotherapeutics Inc.. Lipid-based formulations
US6878805B2 (en) 2002-08-16 2005-04-12 Isis Pharmaceuticals, Inc. Peptide-conjugated oligomeric compounds
US20080039618A1 (en) 2002-11-05 2008-02-14 Charles Allerson Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
US8124745B2 (en) 2002-11-05 2012-02-28 Isis Pharmaceuticals, Inc Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
US7696345B2 (en) 2002-11-05 2010-04-13 Isis Pharmaceuticals, Inc. Polycyclic sugar surrogate-containing oligomeric compounds and compositions for use in gene modulation
WO2004106356A1 (en) 2003-05-27 2004-12-09 Syddansk Universitet Functionalized nucleotide derivatives
US7427672B2 (en) 2003-08-28 2008-09-23 Takeshi Imanishi Artificial nucleic acids of n-o bond crosslinkage type
US20050175682A1 (en) 2003-09-15 2005-08-11 Protiva Biotherapeutics, Inc. Polyethyleneglycol-modified lipid compounds and uses thereof
WO2005071093A2 (en) 2004-01-23 2005-08-04 Istituto Di Ricerche Di Biologia Molecolare P Angeletti Spa Chimpanzee adenovirus vaccine carriers
US20060008378A1 (en) 2004-04-30 2006-01-12 Kunihiro Imai Sterilization method
WO2006048215A1 (en) 2004-11-02 2006-05-11 Istituto Di Ricerche Di Biologia Molecolare P Angeletti Spa Adenoviral amplicon and producer cells for the production of replication-defective adenoviral vectors, methods of preparation and use thereof
US20100062967A1 (en) 2004-12-27 2010-03-11 Silence Therapeutics Ag Coated lipid complexes and their use
US20080020058A1 (en) 2005-02-14 2008-01-24 Sirna Therapeutics, Inc. Lipid nanoparticle based compositions and methods for the delivery of biologically active molecules
US20120011478A1 (en) 2005-02-24 2012-01-12 Texas Instruments Incorporated Merging sub-resolution assist features of a photolithographic mask
US20090012281A1 (en) 2006-01-27 2009-01-08 Isis Pharmaceuticals, Inc. 6-modified bicyclic nucleic acid analogs
US7399845B2 (en) 2006-01-27 2008-07-15 Isis Pharmaceuticals, Inc. 6-modified bicyclic nucleic acid analogs
US8022193B2 (en) 2006-01-27 2011-09-20 Isis Pharmaceuticals, Inc. 6-modified bicyclic nucleic acid analogs
US7569686B1 (en) 2006-01-27 2009-08-04 Isis Pharmaceuticals, Inc. Compounds and methods for synthesis of bicyclic nucleic acid analogs
US7741457B2 (en) 2006-01-27 2010-06-22 Isis Pharmaceuticals, Inc. 6-modified bicyclic nucleic acid analogs
US7547684B2 (en) 2006-05-11 2009-06-16 Isis Pharmaceuticals, Inc. 5′-modified bicyclic nucleic acid analogs
WO2007134181A2 (en) 2006-05-11 2007-11-22 Isis Pharmaceuticals, Inc. 5'-modified bicyclic nucleic acid analogs
US7750131B2 (en) 2006-05-11 2010-07-06 Isis Pharmaceuticals, Inc. 5′-modified bicyclic nucleic acid analogs
US8088746B2 (en) 2006-05-11 2012-01-03 Isis Pharmaceuticals, Inc. Bis-modified bicyclic nucleic acid analogs
US8268980B2 (en) 2006-05-11 2012-09-18 Isis Pharmaceuticals, Inc. 5′-modified bicyclic nucleic acid analogs
US7666854B2 (en) 2006-05-11 2010-02-23 Isis Pharmaceuticals, Inc. Bis-modified bicyclic nucleic acid analogs
US8030467B2 (en) 2006-05-11 2011-10-04 Isis Pharmaceuticals, Inc. 5′-modified bicyclic nucleic acid analogs
US20080042973A1 (en) 2006-07-10 2008-02-21 Memsic, Inc. System for sensing yaw rate using a magnetic field sensor and portable electronic devices using the same
WO2008101157A1 (en) 2007-02-15 2008-08-21 Isis Pharmaceuticals, Inc. 5'-substituted-2'-f modified nucleosides and oligomeric compounds prepared therefrom
US20130011922A1 (en) 2007-03-02 2013-01-10 F/K/A Mdrna, Inc. Nucleic acid compounds for inhibiting gene expression and uses thereof
US8314227B2 (en) 2007-05-22 2012-11-20 Marina Biotech, Inc. Hydroxymethyl substituted RNA oligonucleotides and RNA complexes
US20130096289A1 (en) 2007-05-22 2013-04-18 Marina Biotech, Inc. Hydroxymethyl substituted rna oligonucleotides and rna complexes
US8278425B2 (en) 2007-05-30 2012-10-02 Isis Pharmaceuticals, Inc. N-substituted-aminomethylene bridged bicyclic nucleic acid analogs
US8278426B2 (en) 2007-06-08 2012-10-02 Isis Pharmaceuticals, Inc. Carbocyclic bicyclic nucleic acid analogs
US8278283B2 (en) 2007-07-05 2012-10-02 Isis Pharmaceuticals, Inc. 6-disubstituted or unsaturated bicyclic nucleic acid analogs
US8546556B2 (en) 2007-11-21 2013-10-01 Isis Pharmaceuticals, Inc Carbocyclic alpha-L-bicyclic nucleic acid analogs
US20110117125A1 (en) 2008-01-02 2011-05-19 Tekmira Pharmaceuticals Corporation Compositions and methods for the delivery of nucleic acids
US8530640B2 (en) 2008-02-07 2013-09-10 Isis Pharmaceuticals, Inc. Bicyclic cyclohexitol nucleic acid analogs
WO2009127060A1 (en) 2008-04-15 2009-10-22 Protiva Biotherapeutics, Inc. Novel lipid formulations for nucleic acid delivery
US20100130588A1 (en) 2008-04-15 2010-05-27 Protiva Biotherapeutics, Inc. Novel lipid formulations for nucleic acid delivery
WO2009132131A1 (en) 2008-04-22 2009-10-29 Alnylam Pharmaceuticals, Inc. Amino lipid based improved lipid formulation
WO2009150222A2 (en) 2008-06-12 2009-12-17 Affitech A/S Improved protein expression system
US8501805B2 (en) 2008-09-24 2013-08-06 Isis Pharmaceuticals, Inc. Substituted alpha-L-bicyclic nucleosides
US20110256175A1 (en) 2008-10-09 2011-10-20 The University Of British Columbia Amino lipids and methods for the delivery of nucleic acids
WO2010053572A2 (en) 2008-11-07 2010-05-14 Massachusetts Institute Of Technology Aminoalcohol lipidoids and uses thereof
US20120027796A1 (en) 2008-11-10 2012-02-02 Alnylam Pharmaceuticals, Inc. Novel lipids and compositions for the delivery of therapeutics
US20110313020A1 (en) 2008-12-03 2011-12-22 Marina Biotech, Inc. UsiRNA Complexes
US20110076335A1 (en) 2009-07-01 2011-03-31 Protiva Biotherapeutics, Inc. Novel lipid formulations for delivery of therapeutic agents to solid tumors
US20120202871A1 (en) 2009-07-01 2012-08-09 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
WO2011005861A1 (en) 2009-07-07 2011-01-13 Alnylam Pharmaceuticals, Inc. Oligonucleotide end caps
US9012421B2 (en) 2009-08-06 2015-04-21 Isis Pharmaceuticals, Inc. Bicyclic cyclohexose nucleic acid analogs
US20150191727A1 (en) 2009-08-06 2015-07-09 Isis Pharmaceuticals, Inc. Bicyclic cyclohexose nucleic acid analogs
US20120149894A1 (en) 2009-08-20 2012-06-14 Mark Cameron Novel cationic lipids with various head groups for oligonucleotide delivery
US20130022649A1 (en) 2009-12-01 2013-01-24 Protiva Biotherapeutics, Inc. Snalp formulations containing antioxidants
US20130338210A1 (en) 2009-12-07 2013-12-19 Alnylam Pharmaceuticals, Inc. Compositions for nucleic acid delivery
US20130053572A1 (en) 2010-01-22 2013-02-28 Steven L. Colletti Novel Cationic Lipids for Oligonucleotide Delivery
US20130203836A1 (en) 2010-04-01 2013-08-08 Isis Pharmaceuticals, Inc. 2' and 5' modified monomers and oligonucleotides
US20120287670A1 (en) 2010-04-09 2012-11-15 Sharp Kabushiki Kaisha Light source module and electronic apparatus provided with same
US20130190383A1 (en) 2010-04-26 2013-07-25 Marina Biotech, Inc. Nucleic acid compounds with conformationally restricted monomers and uses thereof
US20130116307A1 (en) 2010-05-12 2013-05-09 Protiva Biotherapeutics Inc. Novel cyclic cationic lipids and methods of use
US20130123338A1 (en) 2010-05-12 2013-05-16 Protiva Biotherapeutics, Inc. Novel cationic lipids and methods of use thereof
US20120027803A1 (en) 2010-06-03 2012-02-02 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US20130090372A1 (en) 2010-06-04 2013-04-11 Brian W. Budzik Novel Low Molecular Weight Cationic Lipids for Oligonucleotide Delivery
US20130303587A1 (en) 2010-06-30 2013-11-14 Protiva Biotherapeutics, Inc. Non-liposomal systems for nucleic acid delivery
US20130323269A1 (en) 2010-07-30 2013-12-05 Muthiah Manoharan Methods and compositions for delivery of active agents
US20130189351A1 (en) 2010-08-31 2013-07-25 Novartis Ag Lipids suitable for liposomal delivery of protein coding rna
US20130178541A1 (en) 2010-09-20 2013-07-11 Matthew G. Stanton Novel low molecular weight cationic lipids for oligonucleotide delivery
US20130274523A1 (en) 2010-09-30 2013-10-17 John A. Bawiec, III Low molecular weight cationic lipids for oligonucleotide delivery
US20130274504A1 (en) 2010-10-21 2013-10-17 Steven L. Colletti Novel Low Molecular Weight Cationic Lipids For Oligonucleotide Delivery
US20170210967A1 (en) 2010-12-06 2017-07-27 Schlumberger Technology Corporation Compositions and methods for well completions
US20140200257A1 (en) 2011-01-11 2014-07-17 Alnylam Pharmaceuticals, Inc. Pegylated lipids and their use for drug delivery
WO2012162210A1 (en) 2011-05-26 2012-11-29 Merck Sharp & Dohme Corp. Ring constrained cationic lipids for oligonucleotide delivery
WO2013016058A1 (en) 2011-07-22 2013-01-31 Merck Sharp & Dohme Corp. Novel bis-nitrogen containing cationic lipids for oligonucleotide delivery
WO2013036868A1 (en) 2011-09-07 2013-03-14 Marina Biotech Inc. Synthesis and uses of nucleic acid compounds with conformationally restricted monomers
US20150203446A1 (en) 2011-09-27 2015-07-23 Takeda Pharmaceutical Company Limited Di-aliphatic substituted pegylated lipids
US10086013B2 (en) 2011-10-27 2018-10-02 Massachusetts Institute Of Technology Amino acid-, peptide- and polypeptide-lipids, isomers, compositions, and uses thereof
WO2013075035A1 (en) 2011-11-18 2013-05-23 Alnylam Pharmaceuticals Rnai agents, compositions and methods of use thereof for treating transthyretin (ttr) associated diseases
US9708628B2 (en) 2011-11-18 2017-07-18 Nof Corporation Cationic lipid having improved intracellular kinetics
US20140308304A1 (en) 2011-12-07 2014-10-16 Alnylam Pharmaceuticals, Inc. Lipids for the delivery of active agents
US20150005363A1 (en) 2011-12-07 2015-01-01 Alnylam Pharmaceuticals, Inc. Branched Alkyl And Cycloalkyl Terminated Biodegradable Lipids For The Delivery Of Active Agents
US20130195920A1 (en) 2011-12-07 2013-08-01 Alnylam Pharmaceuticals, Inc. Biodegradable lipids for the delivery of active agents
US20140039032A1 (en) 2011-12-12 2014-02-06 Kyowa Hakko Kirin Co., Ltd. Lipid nano particles comprising cationic lipid for drug delivery system
WO2013116126A1 (en) 2012-02-01 2013-08-08 Merck Sharp & Dohme Corp. Novel low molecular weight, biodegradable cationic lipids for oligonucleotide delivery
US20150064242A1 (en) 2012-02-24 2015-03-05 Protiva Biotherapeutics, Inc. Trialkyl cationic lipids and methods of use thereof
US20150057373A1 (en) 2012-03-27 2015-02-26 Sirna Therapeutics, Inc DIETHER BASED BIODEGRADABLE CATIONIC LIPIDS FOR siRNA DELIVERY
WO2014062811A2 (en) 2012-10-16 2014-04-24 New York University METHOD OF TREATING CANCER WITH MODULATORS OF SCFSkp2
WO2014136086A1 (en) 2013-03-08 2014-09-12 Novartis Ag Lipids and lipid compositions for the delivery of active agents
US20160151284A1 (en) 2013-07-23 2016-06-02 Protiva Biotherapeutics, Inc. Compositions and methods for delivering messenger rna
WO2015016718A1 (en) 2013-08-02 2015-02-05 Bionovion Holding B.V. Combining cd27 agonists and immune checkpoint inhibition for immune stimulation
US20150140070A1 (en) 2013-10-22 2015-05-21 Shire Human Genetic Therapies, Inc. Lipid formulations for delivery of messenger rna
US20150239926A1 (en) 2013-11-18 2015-08-27 Arcturus Therapeutics, Inc. Asymmetric ionizable cationic lipid for rna delivery
US20150141678A1 (en) 2013-11-18 2015-05-21 Arcturus Therapeutics, Inc. Ionizable cationic lipid for rna delivery
WO2015074085A1 (en) 2013-11-18 2015-05-21 Arcturus Therapeutics, Inc. Ionizable cationic lipid for rna delivery
US20160311759A1 (en) 2013-12-19 2016-10-27 Luis Brito Lipids and Lipid Compositions for the Delivery of Active Agents
US20160317458A1 (en) 2013-12-19 2016-11-03 Luis Brito Lipids and Lipid Compositions for the Delivery of Active Agents
WO2015095340A1 (en) 2013-12-19 2015-06-25 Novartis Ag Lipids and lipid compositions for the delivery of active agents
WO2015106128A2 (en) 2014-01-09 2015-07-16 Alnylam Pharmaceuticals, Inc. MODIFIED RNAi AGENTS
WO2015199952A1 (en) 2014-06-25 2015-12-30 Acuitas Therapeutics Inc. Novel lipids and lipid nanoparticle formulations for delivery of nucleic acids
US20150376115A1 (en) 2014-06-25 2015-12-31 Acuitas Therapeutics Inc. Novel lipids and lipid nanoparticle formulations for delivery of nucleic acids
US20190240349A1 (en) 2015-06-19 2019-08-08 Massachusetts Institute Of Technology Alkenyl substituted 2,5-piperazinediones, compositions, and uses thereof
WO2017004143A1 (en) 2015-06-29 2017-01-05 Acuitas Therapeutics Inc. Lipids and lipid nanoparticle formulations for delivery of nucleic acids
US20160376224A1 (en) 2015-06-29 2016-12-29 Acuitas Therapeutics, Inc. Lipids and lipid nanoparticle formulations for delivery of nucleic acids
US10221127B2 (en) 2015-06-29 2019-03-05 Acuitas Therapeutics, Inc. Lipids and lipid nanoparticle formulations for delivery of nucleic acids
US9867888B2 (en) 2015-09-17 2018-01-16 Modernatx, Inc. Compounds and compositions for intracellular delivery of therapeutic agents
US20170119904A1 (en) 2015-10-28 2017-05-04 Acuitas Therapeutics, Inc. Novel lipids and lipid nanoparticle formulations for delivery of nucleic acids
WO2017099823A1 (en) 2015-12-10 2017-06-15 Modernatx, Inc. Compositions and methods for delivery of therapeutic agents
US20180028664A1 (en) 2015-12-10 2018-02-01 Modernatx, Inc. Compositions and methods for delivery of agents
WO2017117528A1 (en) 2015-12-30 2017-07-06 Acuitas Therapeutics, Inc. Lipids and lipid nanoparticle formulations for delivery of nucleic acids
WO2017173054A1 (en) 2016-03-30 2017-10-05 Intellia Therapeutics, Inc. Lipid nanoparticle formulations for crispr/cas components
US20190136231A1 (en) 2016-03-30 2019-05-09 Intellia Therapeutics, Inc. Lipid nanoparticle formulations for crispr/cas components
WO2018081480A1 (en) 2016-10-26 2018-05-03 Acuitas Therapeutics, Inc. Lipid nanoparticle formulations
WO2019051289A1 (en) 2017-09-08 2019-03-14 Generation Bio Co. Lipid nanoparticle formulations of non-viral, capsid-free dna vectors
WO2019051289A9 (en) 2017-09-08 2019-05-23 Generation Bio Co. Lipid nanoparticle formulations of non-viral, capsid-free dna vectors
WO2019067992A1 (en) 2017-09-29 2019-04-04 Intellia Therapeutics, Inc. Formulations
WO2019157531A1 (en) 2018-02-12 2019-08-15 Ionis Pharmaceuticals, Inc. Modified compounds and uses thereof
WO2019217941A1 (en) 2018-05-11 2019-11-14 Beam Therapeutics Inc. Methods of suppressing pathogenic mutations using programmable base editor systems
WO2020081938A1 (en) 2018-10-18 2020-04-23 Acuitas Therapeutics, Inc. Lipids for lipid nanoparticle delivery of active agents
WO2020106946A1 (en) 2018-11-21 2020-05-28 Translate Bio, Inc. TREATMENT OF CYSTIC FIBROSIS BY DELIVERY OF NEBULIZED mRNA ENCODING CFTR
US11446344B1 (en) 2018-12-12 2022-09-20 Flagship Pioneering Innovations V, Inc. Anellovirus compositions and methods of use
WO2020219876A1 (en) 2019-04-25 2020-10-29 Intellia Therapeutics, Inc. Ionizable amine lipids and lipid nanoparticles
WO2021113777A2 (en) 2019-12-04 2021-06-10 Orna Therapeutics, Inc. Circular rna compositions and methods
WO2021257782A1 (en) 2020-06-18 2021-12-23 Alnylam Pharmaceuticals, Inc. XANTHINE DEHYDROGENASE (XDH) iRNA COMPOSITIONS AND METHODS OF USE THEREOF
WO2022246251A2 (en) 2021-05-21 2022-11-24 Ionis Pharmaceuticals, Inc. Compounds for modulating unc13a expression
WO2022271573A1 (en) 2021-06-21 2022-12-29 Arrowhead Pharmaceuticals, Inc. Rnai agents for inhibiting expression of xanthine dehydrogenase (xdh), pharmaceutical compositions thereof, and methods of use
WO2024049971A2 (en) 2022-09-02 2024-03-07 Baylor College Of Medicine Modulation of b-cell translocation gene 1 (btg1) for use in adoptive cell therapy

Non-Patent Citations (40)

* Cited by examiner, † Cited by third party
Title
"Antisense Drug Technology", 2008, CRC PRESS, pages: 163 - 166,442-443
"The Concise Encyclopedia Of Polymer Science And Engineering", 1990, MACK PUBLISHING CO., pages: 858 - 859
ADLAKHA HUTCHEON ET AL., NAT BIOTECHNOL, vol. 17, 1999, pages 775 - 779
ALBAEK ET AL., J. ORG. CHEM., vol. 71, 2006, pages 7731 - 7740
ALI, NASIR ET AL.: "Current Nucleic Acid Extraction Methods and Their Implications to Point-of-Care Diagnostics", BIOMED RESEARCH INTERNATIONAL, vol. 2017, 2017, pages 9306564
ALTSCHUL SF ET AL., J MOL BIOL, vol. 215, 1990, pages 403
ALTSCHUL SF ET AL., NUC ACIDS RES, vol. 25, 1997, pages 3389 - 3402
COLLOTTA, D ET AL.: "Antisense oligonucleotides: a novel Frontier in pharmacological strategy.", FRONTIERS IN PHARMACOLOGY, vol. 14, 17 November 2023 (2023-11-17), pages 1304342
DONG YSIEGWART DJANDERSON DG.: "Strategies, design, and chemistry in siRNA delivery systems", ADV DRUG DELIV REV, vol. 144, April 2019 (2019-04-01), pages 133 - 147, XP085816831, DOI: 10.1016/j.addr.2019.05.004
DOU, YANMEI ET AL.: "Detecting Somatic Mutations in Normal Cells.", TRENDS IN GENETICS: TIG, vol. 34, no. 7, 2018, pages 545 - 557, XP055729868, DOI: 10.1016/j.tig.2018.04.003
EYGERIS ET AL., NANO LETT., vol. 20, no. 6, 2020, pages 4543 - 4549
FLUITER ET AL., MOL. BIOSYST., vol. 10, 2009, pages 1039
FREIER ET AL., NUCLEIC ACIDS RESEARCH, vol. 25, no. 22, 1997, pages 4429 - 4443
GABIZON ET AL., J CONTROLLED RELEASE, vol. 53, 1998, pages 275 - 279
HOU, X. ET AL., NAT REV MATER, vol. 6, 2021, pages 1078 - 1094, Retrieved from the Internet <URL:https://doi.org/10.1038/s41578-021-00358-0>
HUANG AUGUST YUELEE EUNJUNG ALICE: "Identification of Somatic Mutations From Bulk and Single-Cell Sequencing Data", FRONTIERS IN AGING, vol. 2, 2022
KARLIN SALTSCHUL SF, PNAS, vol. 87, 1990, pages 2264 - 2268
KARLIN SALTSCHUL SF, PNAS, vol. 90, 1993, pages 5873 - 5877
KIM, SANG HYEON ET AL.: "Emerging role of anti-proliferative protein BTG1 and BTG2.", BMB REPORTS, vol. 55, no. 8, 2022, pages 380 - 388
KONING ET AL., BIOCHIM BIOPHYS ACTA, vol. 1420, 1999, pages 153 - 167
KOSHKIN ET AL., TETRAHEDRON, vol. 54, 1998, pages 3607 - 3630
KUMAR ET AL., BIOORG. MED. CHEM. LETT., vol. 8, 1998, pages 2219 - 2222
MARTIN ET AL., CURRENT PHARMACEUTICAL DESIGN, pages 1 - 394
MARTIN ET AL., HELV. CHIM. ACTA, vol. 78, 1995, pages 486 - 504
MARTINCORENA, I. ET AL., CELL, vol. 171, 2017, pages 1029 - 1041
MUYAS, F.SAUER, C.M.VALLE-INCLAN, J.E. ET AL.: "De novo detection of somatic mutations in high-throughput single-cell profiling data sets", NAT BIOTECHNOL, 2023, Retrieved from the Internet <URL:https://doi.org/10.1038/s41587-023-01863-z>
MYERSMILLER, CABIOS, vol. 4, 1988, pages 11 - 17
NIELSEN ET AL., SCIENCE, vol. 254, 1991, pages 1497 - 1500
no. 329918-37-8
NUC. ACIDS SYMP. SERIES, vol. 52, 2008, pages 133 - 134
PARDOLL, NAT REV CANCER, vol. 12, no. 4, 2012, pages 252 - 64
PRAKASH ET AL., ORG. LETT., vol. 5, 2003, pages 403 - 6
RHEINBAY, E. ET AL., NATURE, vol. 578, 2020, pages 102 - 111
SANGHVI, Y.S.: "Antisense Research and Applications", 1993, CRC PRESS, pages: 276 - 278
SHARP ET AL., GENES DEV, vol. 15, 2001, pages 485
SINGH ET AL., CHEM. COMMUN., vol. 4, 1998, pages 455 - 456
SINGH ET AL., J. ORG. CHEM., vol. 63, 1998, pages 10035 - 10039
SRIVASTAVA ET AL., J. AM. CHEM. SOC., vol. 129, 2007, pages 8362 - 8379
TAN, SIUN CHEEBEOW CHIN YIAP.: "DNA, RNA, and protein extraction: the past and the present.", JOURNAL OF BIOMEDICINE & BIOTECHNOLOGY, vol. 2009, 2009, pages 574398, XP002677179, DOI: 10.1155/2009/574398
WORKING ET AL., J PHARMACOL EXP THER, vol. 289, 1999, pages 1128 - 1133

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