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WO2024216191A1 - Modified trems, compositions, and related methods thereof - Google Patents

Modified trems, compositions, and related methods thereof Download PDF

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Publication number
WO2024216191A1
WO2024216191A1 PCT/US2024/024473 US2024024473W WO2024216191A1 WO 2024216191 A1 WO2024216191 A1 WO 2024216191A1 US 2024024473 W US2024024473 W US 2024024473W WO 2024216191 A1 WO2024216191 A1 WO 2024216191A1
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Prior art keywords
trem
alkyl
heteroalkyl
independently
haloalkyl
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PCT/US2024/024473
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French (fr)
Inventor
Theonie ANASTASSIADIS
William F. Kiesman
Hongchuan YU
Daniel Abraham GLAZIER
Christopher E. Arcadia
Qingyi Li
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Flagship Pioneering Innovations VI Inc
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Flagship Pioneering Innovations VI Inc
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Priority to AU2024255972A priority Critical patent/AU2024255972A1/en
Publication of WO2024216191A1 publication Critical patent/WO2024216191A1/en
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

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    • 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
    • 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
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/319—Chemical structure of the backbone linked by 2'-5' linkages, i.e. having a free 3'-position
    • 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
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/323—Chemical structure of the sugar modified ring structure
    • C12N2310/3231—Chemical structure of the sugar modified ring structure having an additional ring, e.g. LNA, ENA

Definitions

  • TREMs modified tRNA-based effector molecules
  • TREMs are complex molecules which can mediate a variety of cellular processes.
  • the TREMs disclosed herein comprise a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, e.g., on a component nucleotide (e.g., a nucleotide sugar).
  • a TREM comprising a sequence of Formula (A): [L1]x-[ASt Domain1]-[L2]x- [DH Domain]-[L3]x-[ACH Domain]-[VL Domain]-[TH Domain]-[L4]x-[ASt Domain2]-[L5]x, wherein independently, [L1] and [VL Domain], are optional; x, independently for each occurrence, is 0 or 1; and one or more of [L1], [ASt Domain1], [L2]-[DH Domain], [L3], [ACH Domain], [VL Domain], [TH Domain], [L4], [ASt Domain2], and [L5] comprises a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide.
  • the TREM comprises a sequence of Formula (A-1): [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]- [ACH Domain]-[VL Domain]-[TH Domain]-[L4]-[ASt Domain2], wherein independently, [L1] and [VL Domain], are optional; wherein one of [L1], [ASt Domain1], [L2]-[DH Domain], [L3], [ACH Domain], [VL Domain], [TH Domain], [L4], and [ASt Domain2] comprises a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide.
  • the TREM further comprises an additional non-naturally occurring modification.
  • the TREMs disclosed herein further comprise at least one additional modification Attorney Docket No.: F2099-7037WO (e.g., a non-naturally occurring modification), e.g., on a component nucleotide (e.g., a nucleobase or sugar) or within an internucleotide region, e.g., the TREM backbone.
  • the TREM comprises feature (a)(i). In an embodiment, the TREM comprises feature (a)(ii). In an embodiment, the TREM comprises feature (a)(iii). In an embodiment, the TREM comprises feature (a)(iv). In an embodiment, the TREM comprises feature (a)(v). In an embodiment, the TREM comprises feature (a)(vi). In an embodiment, the TREM comprises feature (b). In an embodiment, the TREM comprises feature (c). In an embodiment, the TREM comprises feature (d). In an embodiment, the TREM comprises feature (e). In an embodiment, the TREM comprises feature (f). In an embodiment, the TREM comprises all of features (a)-(f) or a combination thereof.
  • the TREM Domain comprising the non-naturally occurring modification has a function, e.g., a domain function described herein.
  • the TREM has the ability to support protein synthesis. In an embodiment, the TREM has the ability to be able to be charged by a synthetase. In an embodiment, the TREM has the ability to be bound by an elongation factor. In an embodiment, the TREM has the ability to introduce an amino acid into a peptide chain. In an embodiment, the TREM has the ability to support elongation. In an embodiment, the TREM has the ability to support initiation. In an embodiment, the [ASt Domain 1] and/or [ASt Domain 2] comprising a non- naturally occurring modification has the ability to initiate or elongate a polypeptide chain.
  • the [ACH Domain] comprising a non-naturally occurring modification has the ability to mediate pairing with a codon.
  • y 1 for any one, two, three, four, five, six, all or a combination of [L1], [L2], [DH Domain], [L3], [VL Domain], [TH Domain], [L4].
  • y 0 for any one, two, three, four, five, six, all or a combination of [L1], [L2], [DH Domain], [L3], [VL Domain], [TH Domain], [L4].
  • y 1 for linker [L1], and L1 comprises a nucleotide having a non- naturally occurring modification.
  • y 1 for linker [L2], and L2 comprises a nucleotide having a non- naturally occurring modification.
  • y 1 for [DH Domain (DHD)], and DHD comprises a nucleotide having a non-naturally occurring modification.
  • the DHD comprising the non- naturally occurring modification has the ability to mediate recognition of aminoacyl-tRNA synthetase.
  • y 1 for linker [L3], and L3 comprises a nucleotide having a non- naturally occurring modification.
  • y 1 for [VL Domain (VLD)], and VLD comprises a nucleotide having a non-naturally occurring modification.
  • y 1 for [TH Domain (THD)], and THD comprises a nucleotide having a non-naturally occurring modification.
  • THD comprising the non- naturally occurring modification has the ability to mediate recognition of the ribosome.
  • y 1 for linker [L4], and L4 comprises a nucleotide having a non- naturally occurring modification.
  • the disclosure provides a TREM fragment comprising a portion of a TREM, wherein the TREM comprises a sequence of Formula (A-1): [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2], and wherein the TREM fragment comprises a non-naturally occurring modification.
  • the TREM fragment comprises one, two, three or all or any combination of the following: (a) a TREM half (e.g., from a cleavage in the ACH Domain, e.g., in the anticodon sequence, e.g., a 5’half or a 3’ half); (b) a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DH Domain or the ACH Domain); (c) a 3’ fragment (e.g., a fragment comprising the 3’ end, e.g., from a cleavage in the TH Domain); or (d) an internal fragment (e.g., from a cleavage in any one of the ACH Domain, DH Domain or TH Domain).
  • a TREM half e.g., from a cleavage in the ACH Domain, e.g., in the anticodon sequence, e.g., a 5
  • the TREM fragment comprise (a) a TREM half which comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the TREM fragment comprise (b) a 5’ fragment which comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the TREM fragment comprise (c) a 3’ fragment which comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the TREM fragment comprise (d) an internal fragment which comprises a nucleotide having a non-naturally occurring modification.
  • the TREM Domain comprises a plurality of nucleotides each having a non- naturally occurring modification.
  • the non-naturally occurring modification comprises a nucleobase modification, a sugar (e.g., ribose) modification, or a backbone modification.
  • tbe non-naturally occurring modification is a sugar (e.g., ribose) modification.
  • tbe non-naturally occurring modification is 2’-ribose Attorney Docket No.: F2099-7037WO modification, e.g., a 2’-OMe, 2’-halo (e.g., 2’-F), 2’-MOE, or 2’-deoxy modification.
  • tbe non-naturally occurring modification is a backbone modification, e.g., a phosphorothioate modification.
  • the TREM sequence comprises a CCA sequence on a terminus, e.g., the 3’ terminus.
  • the TREM sequence does not comprise a CCA sequence on a terminus, e.g., the 3’ terminus.
  • the non-naturally occurring modification is a base modification chosen from a modification listed in Table 5.
  • the TREM, TREM core fragment, or TREM fragments disclosed herein is encoded by a sequence provided in Table 1, e.g., any one of SEQ ID NOs 1-451.
  • the TREM, TREM core fragment, or TREM fragment is encoded by a consensus sequence chosen from any one of SEQ ID NOs: 562-621.
  • the TREM, TREM core fragment, or TREM fragment is encoded by a sequence provided in FIG.1, e.g., any one of SEQ ID NOs: 625-1416, e.g., SEQ ID NOs: 625-793, 794- 991, or 992-1416.
  • the TREM, TREM core fragment, or TREM fragment comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in FIG.1, e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416.
  • the TREM, TREM core fragment, or TREM fragment comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from a TREM provided in FIG.1, e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416.
  • the TREM, TREM core fragment, or TREM fragment comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non- naturally occurring modifications compared with a TREM, TREM core fragment, or TREM fragment provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications), e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416.
  • 2’-ribose modifications or an internucleotide modification e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications
  • SEQ ID Nos: 625-1416 e.g., any one of SEQ ID NOs: 625-7
  • the TREM, TREM core fragment, or TREM fragment comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional LNA moieties or 2’,5’-linked nucleotides compared with a TREM, TREM core fragment, or TREM fragment provided in FIG.1, e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416.
  • the TREM, TREM core fragment, or TREM fragment is a TREM provided in FIG.1, e.g., any one of TREM NOs: 1-791, e.g., any one of TREM NOs: 1-169, 170-367, or 368-791.
  • the TREM, TREM core fragment, or TREM fragment comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications), e.g., any one of TREM NOs: 1-791, e.g., any one of TREM Nos: 1-169, 170-367, or 368-791.
  • the disclosure provides a pharmaceutical composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein.
  • a TREM or a related composition thereof can be used, inter alia, to modulate a functional parameter (e.g., an expression parameter and/or a signaling parameter) of an RNA corresponding to, or a polypeptide encoded by, a nucleic acid sequence comprising an endogenous open reading frame (ORF) having a premature termination codon (PTC).
  • a functional parameter e.g., an expression parameter and/or a signaling parameter
  • ORF endogenous open reading frame
  • PTC premature termination codon
  • a method of modulating a functional parameter of an mRNA corresponding to, or polypeptide encoded by, an endogenous open reading frame (ORF) in a subject which ORF comprises a premature termination codon (PTC)
  • ORF an endogenous open reading frame
  • PTC premature termination codon
  • contacting the subject with a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein (e.g., a TREM, TREM core fragment, or a TREM fragment comprising an LNA moiety or a 2’,5’-linked nucleotide) in an amount and/or for a time sufficient to modulate the functional parameter of the mRNA or polypeptide, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the codon having the first sequence, thereby modulating the functional parameter in the subject.
  • a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein (
  • the functional parameter comprises a signaling parameter and/or an expression parameter, e.g., as described herein.
  • a method of modulating expression of a protein in a cell wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), which ORF comprises a premature termination codon (PTC), comprising Attorney Docket No.: F2099-7037WO contacting the cell with a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein (e.g., a TREM, TREM core fragment, or a TREM fragment comprising an LNA moiety or a 2’,5’-linked nucleotide) in an amount and/or for a time sufficient to modulate expression of the encoded protein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the PTC, thereby modulating expression of the protein in the cell.
  • ORF endogenous open reading frame
  • PTC premature termination
  • the PTC comprises UAA, UGA or UAG.
  • a method of increasing expression of a protein in a subject wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), which ORF comprises a premature termination codon (PTC), comprising contacting the subject, in an amount and/or for a time sufficient to increase expression of the protein, with a TREM composition that (i) has an anticodon that pairs with the PTC, (ii) recognizes an aminoacyl-tRNA synthetase specific for Trp, Tyr, Cys, Glu, Lys, Gln, Ser, Leu, Arg, or Gly, (iii) comprises a sequence of Formula A, or (iv) comprises a non-naturally occurring modification.
  • ORF endogenous open reading frame
  • PTC premature termination codon
  • the PTC comprises UAA, UGA or UAG.
  • the TREM composition comprises (i). In an embodiment, the TREM composition comprises (ii). In an embodiment, the TREM composition comprises (iii). In an embodiment, the TREM composition comprises (iv). In an embodiment, the TREM composition comprises two of (i)- (iv). In an embodiment, the TREM composition comprises three of (i)-(iv). In an embodiment, the TREM composition comprises each of (i)-(iv).
  • the disclosure provides a method of treating a subject having an endogenous open reading frame (ORF) which comprises a premature termination codon (PTC), comprising providing a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM comprises an anticodon that pairs with the PTC in the ORF; contacting the subject with the composition comprising a TREM, TREM core fragment or TREM fragment in an amount and/or for a time sufficient to treat the subject, thereby treating the subject.
  • the PTC comprises UAA, UGA or UAG.
  • the disclosure provides a method of treating a subject having an disease or disorder associated with a premature termination codon (PTC), comprising providing a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein (e.g., a TREM, TREM core fragment, or a TREM fragment comprising an LNA moiety or a 2’,5’-linked nucleotide); contacting the subject with the composition comprising a TREM, Attorney Docket No.: F2099-7037WO TREM core fragment or TREM fragment in an amount and/or for a time sufficient to treat the subject, thereby treating the subject.
  • the PTC comprises UAA, UGA or UAG.
  • the disease or disorder associated with a PTC is a disease or disorcer described herein, e.g., a cancer or a monogenic disease.
  • the codon having the first sequence comprises a mutation (e.g., a point mutation, e.g., a nonsense mutation), resulting in a premature termination codon (PTC) chosen from UAA, UGA or UAG.
  • the codon having the first sequence or the PTC comprises a UAA mutation.
  • the codon having the first sequence or the PTC comprises a UGA mutation.
  • the codon having the first sequence or the PTC comprises a UAG mutation
  • the disclosure provides a method of making a TREM, a TREM core fragment, or a TREM fragment disclosed herein, comprising linking a first nucleotide to a second nucleotide to form the TREM.
  • the TREM, TREM core fragment or TREM fragment is non-naturally occurring (e.g., synthetic).
  • the TREM, TREM core fragment or TREM fragment is made by cell- free solid phase synthesis.
  • the disclosure provides a method of modulating a tRNA pool in a cell comprising: providing a TREM, a TREM core fragment, or a TREM fragment disclosed herein, and contacting the cell with the TREM, TREM core fragment or TREM fragment, thereby modulating the tRNA pool in the cell.
  • the disclosure provides a method of contacting a cell, tissue, or subject with a TREM, a TREM core fragment, or a TREM fragment disclosed herein, comprising: contacting the cell, tissue or subject with the TREM, TREM core fragment or TREM fragment, thereby contacting the cell, tissue, or subject with the TREM, TREM core fragment or TREM fragment.
  • the disclosure provides a method of delivering a TREM, TREM core fragment or TREM fragment to a cell, tissue, or subject, comprising: providing a cell, tissue, or subject, and contacting the cell, tissue, or subject, a TREM, a TREM core fragment, or a TREM fragment disclosed herein.
  • the disclosure provides a method of modulating a tRNA pool in a cell comprising an endogenous open reading frame (ORF), which ORF comprises a codon having a first sequence, comprising: optionally, acquiring knowledge of the abundance of one or both of (i) and (ii), e.g., acquiring knowledge of the relative amounts of: (i) and (ii) in the cell, wherein (i) is a tRNA moiety having an anticodon that pairs with the codon of the ORF having a first sequence (the first tRNA moiety) and (ii) is an isoacceptor tRNA moiety having an anticodon that pairs with a codon other than the codon having the first sequence (the second tRNA moiety) in the cell; contacting the cell with a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that
  • the disclosure provides a method of modulating a tRNA pool in a subject having an ORF, which ORF comprises a codon having a first sequence, comprising: optionally, acquiring knowledge of the abundance of one or both of (i) and (ii), e.g., acquiring knowledge of the relative amounts of: (i) and (ii) in the subject, wherein (i) is a tRNA moiety having an anticodon that pairs with the codon of the ORF having a first sequence (the first tRNA moiety) and (ii) is an isoacceptor tRNA moiety having an anticodon that pairs with a codon other than the codon having the first sequence (the second tRNA moiety) in the subject; contacting the subject with a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with: the codon having the first sequence; or the codon other than the codon having the
  • the disclosure provides a method of modulating a tRNA pool in a subject having an endogenous ORF comprising a codon comprising a synonymous mutation (a synonymous mutation codon or SMC), comprising: Attorney Docket No.: F2099-7037WO providing a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment comprises an isoacceptor tRNA moiety comprising an anticodon sequence that pairs with the SMC (the TREM); contacting the subject with the composition in an amount and/or for a time sufficient to modulate the tRNA pool in the subject, thereby modulating the tRNA pool in the subject.
  • a synonymous mutation a synonymous mutation codon or SMC
  • the disclosure provides a method of modulating a tRNA pool in a cell comprising an endogenous ORF comprising a codon comprising a SMC, comprising: providing a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment comprises an isoacceptor tRNA moiety comprising an anticodon sequence that pairs with the SMC (the TREM); contacting the cell with the composition comprising a TREM in an amount and/or for a time sufficient to modulate the tRNA pool in the cell, thereby modulating the tRNA pool in the cell.
  • the disclosure provides a method of modulating expression of a protein in a cell, wherein the protein is encoded by a nucleic acid comprising an ORF, which ORF comprises a codon having a mutation, comprising: contacting the cell with a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein in an amount and/or for a time sufficient to modulate expression of the encoded protein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the codon having the mutation, thereby modulating expression of the protein in the cell.
  • the disclosure provides a method of modulating expression of a protein in a subject, wherein the protein is encoded by a nucleic acid comprising an endogenous ORF, which ORF comprises a codon having a mutation, comprising: contacting the subject with a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, in an amount and/or for a time sufficient to modulate expression of the encoded protein, Attorney Docket No.: F2099-7037WO wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the codon having the mutation, thereby modulating expression of the protein in the subject.
  • the mutation in the ORF is a nonsense mutation, e.g., resulting in a premature stop codon chosen from UAA, UGA or UAG.
  • the stop codon is UAA.
  • the stop codon is UGA.
  • the stop codon is UAG.
  • the TREM comprises an anticodon that pairs with a stop codon.
  • TREMs of the disclosure include TREMs, TREM core fragments and TREM fragments. TREMs, TREM core fragments or TREM fragments can be modified with non-naturally occurring modifications to, e.g., increase the level and/or activity (e.g., stability) of the TREM.
  • TREM compositions e.g., comprising TREMs having a non-naturally occurring modification
  • TREMs can be administered to cells, tissues or subjects to modulate these functions, e.g., in vitro or in vivo.
  • TREMs, TREM core fragments or TREM fragments comprising non-naturally occurring modifications
  • TREM compositions, preparations, methods of making TREM compositions and preparations, and methods of using the same e.g., the TREM, TREM core fragment, and TREM fragments comprise a non-naturally occurring modification that improves stability or enhances activity of the TREM, TREM core fragment, or TREM fragment.
  • TREMs any of the aforesaid TREMs, TREM core fragments, TREM fragments, TREM compositions, preparations, methods of making TREM compositions and preparations, and methods of using TREM compositions and preparations
  • TREMs TREM core fragments, TREM fragments, TREM compositions, preparations, methods of making TREM compositions and preparations, and methods of using TREM compositions and preparations
  • Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following Enumerated Embodiments, Drawings, Description, Examples, or Claims.
  • FIG.1 is a listing of exemplary TREMs described herein, e.g., TREMs comprising a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide.
  • LNA locked nucleic acid
  • FIG.1 is a listing of exemplary TREMs described herein, e.g., TREMs comprising a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide.
  • LNA locked nucleic acid
  • CERTAIN EMBODIMENTS The present disclosure features tRNA-based effector molecules (TREMs) comprising a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, as well as compositions and related methods of use and making.
  • TREMs tRNA-based effector molecules
  • TREMs are complex molecules which can mediate a variety of cellular processes.
  • Pharmaceutical TREM compositions e.g., TREMs comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, can be administered to a cell, a tissue, or to a subject to modulate these functions.
  • ORF endogenous open reading frame
  • TREMs comprising a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide, and an additional non-naturally occurring modification, methods of making the same and compositions thereof.
  • a “disease or disorder associated with a PTC” as that term is used herein includes, but is not limited to, a disease or disorder in which cells express, or at one time expressed, a polypeptide encoded by an ORF comprising a PTC.
  • a disease associated with a PTC is chosen from: a proliferative disorder (e.g., a cancer), a genetic disorder, a metabolic disorder, an immune disorder, an inflammatory disorder or a neurological disorder.
  • exemplary diseases or disorders associated with a PTC are provided in any one of Tables 15, 16 Attorney Docket No.: F2099-7037WO and 17.
  • the disease associated with a PTC is a cancer.
  • the disease associated with a PTC is a monogenic disease.
  • an “isoacceptor,” as that term is used herein, refers to a plurality of tRNA molecule or TREMs wherein each molecule of the plurality comprises a different naturally occurring anticodon sequence and each molecule of the plurality mediates the incorporation of the same amino acid and that amino acid is the amino acid that naturally corresponds to the anticodons of the plurality.
  • a “modification,” as that term is used herein with reference to a nucleotide refers to a modification of the chemical structure, e.g., a covalent modification, of the subject nucleotide. The modification can be naturally occurring or non-naturally occurring. In an embodiment, the modification is non-naturally occurring. In an embodiment, the modification is naturally occurring.
  • the modification is a synthetic modification. In an embodiment, the modification is a modification provided in Table 5.
  • a “naturally occurring nucleotide,” as that term is used herein, refers to a nucleotide that does not comprise a non-naturally occurring modification. In an embodiment, it includes a naturally occurring modification.
  • the modification is added synthetically, e.g., in a cell free reaction, e.g., in a solid state or liquid phase synthetic reaction.
  • the non-naturally occurring modification is a modification that is not present (in identity, location or position) if a sequence of the TREM is expressed in a mammalian cell, e.g., a HEK293 cell line. Exemplary non-naturally occurring modifications are found in Table 5.
  • the non-naturally occurring modification comprises a locked nucleic acid moiety or a 2’,5’-linked nucleotide.
  • the non- naturally occurring modification comprises a locked nucleic acid moiety.
  • the non-naturally occurring modification comprises a 2’,5’-linked nucleotide.
  • Attorney Docket No.: F2099-7037WO A “non-naturally modified nucleotide,” as that term is used herein, refers a nucleotide comprising a non-naturally occurring modification on or of a sugar, nucleobase, or phosphate moiety.
  • a nucleotide comprises a naturally occurring, e.g., naturally occurring in a human cell, nucleotide, e.g., an adenine, thymine, guanine, cytosine, or uracil nucleotide.
  • ORF open reading frame
  • a PTC occurs at a position upstream of a naturally occurring stop codon in an ORF.
  • a PTC can differ (or arise) from a pre-mutation sequence by a point mutation, e.g., a nonsense mutation.
  • a PTC can differ (or arise) from a pre-mutation sequence by a genetic change, e.g., abnormality, other than a point mutation, e.g., a frameshift, a deletion, an insertion, a rearrangement, an inversion, a translocation, a duplication, or a transversion.
  • a PTC results in the production of a truncated protein which lacks a native activity or which is associated with a mutant, disease, or other unwanted phenotype.
  • the ORF comprising the PTC is an ORF from a tumor suppressor gene.
  • the mutation giving rise to the PTC is a driver mutation, e.g., a mutation that provides a growth advantage to a tumor cell.
  • a “functional parameter,” refers to an expression parameter and/or a signaling parameter.
  • a functional parameter is an expression parameter.
  • An expression parameter includes an expression parameter of a polypeptide or protein encoded by the endogenous ORF having a first sequence or PTC; or an expression parameter of an RNA, e.g., messenger RNA, encoded by the endogenous ORF having a first sequence or PTC.
  • an expression parameter can include: (a) protein translation; (b) expression level (e.g., of polypeptide or protein, or mRNA); (c) post-translational modification of polypeptide or protein; (d) folding (e.g., of polypeptide or protein, or mRNA), Attorney Docket No.: F2099-7037WO (e) structure (e.g., of polypeptide or protein, or mRNA), (f) transduction (e.g., of polypeptide or protein), (g) compartmentalization (e.g., of polypeptide or protein, or mRNA), (h) incorporation (e.g., of polypeptide or protein, or mRNA) into a supermolecular structure, e.g., incorporation into a membrane, proteasome, or ribosome, (i) incorporation into a multimeric polypeptide, e.g., a homo or heterodimer, and/or (j) stability.
  • e protein translation
  • expression level
  • a functional parameter is a signaling parameter.
  • a signaling parameter can include: (1) modulation of a signaling pathway, e.g., a cellular signaling pathway which is downstream or upstream of the protein encoded by the endogenous ORF having a first sequence or PTC; (2) cell fate modulation; (3) ribosome occupancy modulation; (4) protein translation modulation; (5) mRNA stability modulation; (6) protein folding and structure modulation; (7) protein transduction or compartmentalization modulation; and/or (8) protein stability modulation.
  • the ORF having the PTC is associated with a disease or disorder associated with a PTC, e.g., as described herein, e.g., a disease or disorder listed in any one of Tables 15, 16 and 17. In an embodiment, the ORF having the PTC is not associated with a disease or disorder associated with a PTC.
  • a “stop codon” as that term is used herein, refers to a three nucleotide contiguous sequence within messenger RNA that specifies a termination of translation. For example, UAG, UAA, UGA (in RNA) and TAG, TAA or TGA (in DNA) are stop codons.
  • tRNA-based effector molecule refers to an RNA molecule comprising a structure or property from (a)-(v) below, and which is a recombinant TREM, a synthetic TREM, or a TREM expressed from a heterologous cell.
  • the Attorney Docket No.: F2099-7037WO TREMs described in the present disclosure comprise a nucleotide bearing a locked nucleic acid moiety or a 2’,5’-linked nucleotide.
  • TREMs described herein are synthetic molecules and are made, e.g., in a cell free reaction, e.g., in a solid state or liquid phase synthetic reaction.
  • TREMs are chemically distinct, e.g., in terms of primary sequence, type or location of modifications from the endogenous tRNA molecules made in cells, e.g., in mammalian cells, e.g., in human cells.
  • a TREM can have a plurality (e.g., 2, 3, 4, 5, 6, 7, 8, 9) of the structures and functions of (a)-(v).
  • a TREM is non-native, as evaluated by structure or the way in which it was made.
  • a TREM comprises one or more of the following structures or properties: (a’) an optional linker region of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 1 region; (a) an amino acid attachment domain that binds an amino acid, e.g., an acceptor stem domain (AStD), wherein an AStD comprises sufficient RNA sequence to mediate, e.g., when present in an otherwise wildtype tRNA, acceptance of an amino acid, e.g., its cognate amino acid or a non-cognate amino acid, and transfer of the amino acid (AA) in the initiation or elongation of a polypeptide chain.
  • AStD acceptor stem domain
  • the AStD comprises a 3’-end adenosine (CCA) for acceptor stem charging which is part of synthetase recognition.
  • CCA 3’-end adenosine
  • the AStD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring AStD, e.g., an AStD encoded by a nucleic acid in Table 1.
  • the TREM can comprise a fragment or analog of an AStD, e.g., an AStD encoded by a nucleic acid in Table 1, which fragment in embodiments has AStD activity and in other embodiments does not have AStD activity.
  • AStD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions; Attorney Docket No.: F2099-7037WO
  • AStD comprises residues R 1 -R 2 -R 3 -R 4 -R 5 -R 6 -R 7 and residues R 65 - R66-R67-R68-R69-R70-R71 of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids;
  • AStD comprises residues R1-R2-R3-R4
  • a DHD mediates the stabilization of the TREM’s tertiary structure.
  • the DHD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring DHD, e.g., a DHD encoded by a nucleic acid in Table 1.
  • the TREM can comprise a fragment or analog of a DHD, e.g., a DHD encoded by a nucleic acid in Table 1, which fragment in embodiments has DHD activity and in other embodiments does not have DHD activity.
  • the DHD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;
  • the DHD comprises residues R10-R11-R12-R13-R14 R15-R16-R17-R18- R19-R20-R21-R22-R23-R24-R25-R26-R27-R28 of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids;
  • the DHD comprises residues R 10 -R 11 -R 12 -R 13 -R 14 R 15 -R 16 -R 17 -R 18 - R19-R20-R21-R22-R23-R24-R25-R26-R27-R28 of Formula II ZZZ, wherein ZZZ indicates any of the twenty amino acids;
  • the DHD comprises residues R 10 -R 11 -R 12 -R 13 -R 14
  • the TREM can comprise a fragment or analog of an ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1, which fragment in embodiments has ACHD activity and in other embodiments does not have ACHD activity.
  • the ACHD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;
  • the ACHD comprises residues -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 - R39-R40-R41-R42-R43-R44-R45-R46 of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids;
  • the ACHD comprises residues -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 - R 39 -R 40
  • a VLD mediates the stabilization of the TREM’s tertiary structure.
  • a VLD modulates, e.g., increases, the specificity of the TREM, e.g., for its cognate amino acid, e.g., the VLD modulates the TREM’s cognate adaptor function.
  • the VLD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring VLD, e.g., a VLD encoded by a nucleic acid in Table 1.
  • the THD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring THD, e.g., a THD encoded by a nucleic acid in Table 1.
  • the TREM can comprise a fragment or analog of a THD, e.g., a THD encoded by a nucleic acid in Table 1, which fragment in embodiments has THD activity and in other embodiments does not have THD activity.
  • the THD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions;
  • the THD comprises residues -R48-R49-R50-R51-R52-R53-R54-R55-R56- R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 of Formula I ZZZ , wherein ZZZ indicates any of the twenty amino acids;
  • the THD comprises residues -R48-R49-R50-R51-R52-R53-R54-R55-R56- R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 of Formula II ZZZ , wherein ZZZ indicates any of the twenty amino acids;
  • a loop can comprise a domain described herein, e.g., a domain selected from (a)-(e).
  • a loop can comprise one or a plurality of domains.
  • a stem or loop structure has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1.
  • the TREM can comprise a fragment or analog of a stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1, which fragment in embodiments has activity of a stem or loop structure, and in other embodiments does not have activity of a stem or loop structure; (g) a tertiary structure, e.g., an L-shaped tertiary structure; (h) adaptor function, i.e., the TREM mediates acceptance of an amino acid, e.g., its cognate amino acid and transfer of the AA in the initiation or elongation of a polypeptide chain; (i) cognate adaptor function wherein the TREM mediates acceptance and incorporation of an amino acid (e.g., cognate amino acid) associated in nature with the anti-codon of the TREM to initiate or elongate a polypeptide chain; (j) non-cognate adaptor function, wherein the TREM mediates acceptance and incorporation of an amino acid (
  • a TREM comprises the following properties: (a), (c), (h), (b), (e) and (g). In an embodiment, a TREM comprises the following properties: (a), (c), (h) and (m). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), and (g). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m) and (b). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m) and (e). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), (g), (b) and (e).
  • a TREM comprises a linker, e.g., an RNA linker, e.g., a flexible RNA linker, which provides for covalent linkage between a first and a second structure or domain.
  • an RNA linker comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 ribonucleotides.
  • a TREM can comprise one or a plurality of linkers, e.g., in embodiments a TREM comprising (a), (b), (c), (d) and (e) can have a first linker between a first and second domain, and a second linker between a third domain and another domain.
  • the TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]- [L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2].
  • a TREM comprises an RNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with, or which differs by no more than 1, 2, 3, 4, 5, 10, 15, 20, 25, or 30 ribonucleotides from, an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof.
  • a TREM comprises a TREM domain, e.g., a domain described herein, comprising at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical with, or which differs by no more than 1, 2, 3, 4, 5, 10, or 15, ribonucleotides from, an RNA encoded by a DNA sequence listed in Table 1, or a fragment or a functional fragment thereof.
  • a TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by DNA sequence listed in Table 1, or a fragment or functional fragment thereof.
  • a TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by DNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with a DNA sequence listed in Table 1, or a fragment or functional fragment thereof.
  • a TREM is 76-90 nucleotides in length.
  • a TREM or a fragment or functional fragment thereof is between 10-90 nucleotides, between 10-80 nucleotides, between 10-70 nucleotides, between 10-60 nucleotides, between 10-50 nucleotides, between 10-40 nucleotides, between 10-30 nucleotides, between 10-20 nucleotides, between 20- 90 nucleotides, between 20-80 nucleotides, 20-70 nucleotides, between 20-60 nucleotides, between 20-50 nucleotides, between 20-40 nucleotides, between 30-90 nucleotides, between 30- 80 nucleotides, between 30-70 nucleotides, between 30-60 nucleotides, or between 30-50 nucleotides.
  • a TREM is aminoacylated, e.g., charged, with an amino acid by an aminoacyl tRNA synthetase. In an embodiment, a TREM is not charged with an amino acid, e.g., an uncharged TREM (uTREM). In an embodiment, a TREM comprises less than a full length tRNA. In embodiments, a TREM can correspond to a naturally occurring fragment of a tRNA, or to a non-naturally occurring fragment.
  • Exemplary fragments include: TREM halves (e.g., from a cleavage in the ACHD, e.g., in the anticodon sequence, e.g., 5’halves or 3’ halves); a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DHD or the ACHD); a 3’ fragment (e.g., a fragment comprising the 3’ end, e.g., from a cleavage in the THD); or an internal fragment (e.g., from a cleavage in one or more of the ACHD, DHD or THD).
  • TREM halves e.g., from a cleavage in the ACHD, e.g., in the anticodon sequence, e.g., 5’halves or 3’ halves
  • a 5’ fragment e.g., a fragment comprising the 5’ end, e.g., from
  • a “TREM fragment,” as used herein, refers to a portion of a TREM, wherein the TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2].
  • “Decreased expression,” as that term is used herein, refers to a decrease in comparison to a reference, e.g., in the case where altered control region, or addition of an agent, results in a Attorney Docket No.: F2099-7037WO decreased expression of the subject product, it is decreased relative to an otherwise similar cell without the alteration or addition.
  • An “exogenous nucleic acid,” as that term is used herein, refers to a nucleic acid sequence that is not present in or differs by at least one nucleotide from the closest sequence in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced.
  • an exogenous nucleic acid comprises a nucleic acid that encodes a TREM.
  • exogenous TREM refers to a TREM that: (a) differs by at least one nucleotide or one post transcriptional modification from the closest sequence tRNA in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced; (b) has been introduced into a cell other than the cell in which it was transcribed; (c) is present in a cell other than one in which it naturally occurs; or (d) has an expression profile, e.g., level or distribution, that is non-wildtype, e.g., it is expressed at a higher level than wildtype.
  • the expression profile can be mediated by a change introduced into a nucleic acid that modulates expression or by addition of an agent that modulates expression of the RNA molecule.
  • an exogenous TREM comprises 1, 2, 3 or 4 of properties (a)-(d).
  • a “GMP-grade composition,” as that term is used herein, refers to a composition in compliance with current good manufacturing practice (cGMP) guidelines, or other similar requirements.
  • cGMP current good manufacturing practice
  • a GMP-grade composition can be used as a pharmaceutical product.
  • the terms “increasing” and “decreasing” refer to modulating that results in, respectively, greater or lesser amounts of function, expression, or activity of a particular metric relative to a reference.
  • the amount of a marker of a metric may be increased or decreased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98%, 2X, 3X, 5X, 10X or more relative to the amount of the marker prior to administration or relative to the effect of a negative control agent.
  • the metric may be measured subsequent to administration at a time that the administration has had the recited effect, e.g., at Attorney Docket No.: F2099-7037WO least 12 hours, 24 hours, one week, one month, 3 months, or 6 months, after a treatment has begun.
  • Increased expression refers to an increase in comparison to a reference, e.g., in the case where altered control region, or addition of an agent, results in an increased expression of the subject product, it is increased relative to an otherwise similar cell without the alteration or addition.
  • non-cognate adaptor function TREM refers to a TREM which mediates initiation or elongation with an AA (a non-cognate AA) other than the AA associated in nature with the anti-codon of the TREM.
  • a non-cognate adaptor function TREM is also referred to as a mischarged TREM (mTREM).
  • a “non-naturally occurring sequence,” as that term is used herein, refers to a sequence wherein an Adenine is replaced by a residue other than an analog of Adenine, a Cytosine is replaced by a residue other than an analog of Cytosine, a Guanine is replaced by a residue other than an analog of Guanine, and a Uracil is replaced by a residue other than an analog of Uracil.
  • An analog refers to any possible derivative of the ribonucleotides, A, G, C or U.
  • a sequence having a derivative of any one of ribonucleotides A, G, C or U is a non- naturally occurring sequence.
  • a “pharmaceutical TREM composition,” as that term is used herein, refers to a TREM composition that is suitable for pharmaceutical use.
  • a pharmaceutical TREM composition comprises a pharmaceutical excipient.
  • the TREM will be the only active ingredient in the pharmaceutical TREM composition.
  • the pharmaceutical TREM composition is free, substantially free, or has less than a pharmaceutically acceptable amount, of host cell proteins, DNA, e.g., host cell DNA, endotoxins, and bacteria.
  • the covalent modification occurs co-transcriptionally.
  • the modification is made in vivo, e.g., in a cell used to produce a TREM.
  • the modification is made ex vivo, e.g., it is made on a TREM isolated or obtained from the cell which produced the TREM.
  • the post-transcriptional modification is selected from a post-transcriptional modification listed in Table 2.
  • a “synthetic TREM,” as that term is used herein, refers to a TREM which was synthesized other than in or by a cell having an endogenous nucleic acid encoding the TREM, e.g., a synthetic TREM is synthetized by cell-free solid phase synthesis.
  • a synthetic TREM can have the same, or a different, sequence, or tertiary structure, as a native tRNA.
  • a “recombinant TREM,” as that term is used herein, refers to a TREM that was expressed in a cell modified by human intervention, having a modification that mediates the production of the TREM, e.g., the cell comprises an exogenous sequence encoding the TREM, or a modification that mediates expression, e.g., transcriptional expression or post-transcriptional modification, of the TREM.
  • a recombinant TREM can have the same, or a different, sequence, set of post-transcriptional modifications, or tertiary structure, as a reference tRNA, e.g., a native tRNA.
  • a TREM composition can comprise one or more species of TREMs, TREM core fragments or TREM fragments. In an embodiment, the composition comprises only a single species of TREM, TREM core fragment or TREM fragment. In an embodiment, the TREM composition comprises a first TREM, TREM core fragment or TREM fragment species; and a second TREM, TREM core fragment or TREM fragment species.
  • the TREM, TREM core fragment or TREM fragment has at least 70, 75, 80, 85, 90, or 95, or has 100%, identity with a sequence encoded by a nucleic acid in Table 1.
  • a TREM composition can comprise one or more species of TREMs, TREM core fragments or TREM fragments.
  • the TREM composition is at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% dry weight TREMs (for a liquid composition dry weight refers to the weight after removal of substantially all liquid, e.g., after lyophilization).
  • the composition is a liquid. In an embodiment, the composition is dry, e.g., a lyophilized material. In an embodiment, the composition is a frozen composition. In an embodiment, the composition is sterile. In an embodiment, the composition comprises at least 0.5 g, 1.0 g, 5.0 g, 10 g, 15 g, 25 g, 50 g, 100 g, 200 g, 400 g, or 500 g (e.g., as determined by dry weight) of TREM.
  • At least X% of the TREMs in a TREM composition has a non- naturally occurring modification at a selected position, and X is 80, 90, 95, 96, 97, 98, 99, or 99.5.
  • at least X% of the TREMs in a TREM composition has a non- naturally occurring modification at a first position and a non-naturally occurring modification at a second position, and X, independently, is 80, 90, 95, 96, 97, 98, 99, or 99.5.
  • the modification at the first and second position is the same. In embodiments, the modification at the first and second position are different.
  • the nucleiotide at the first and second position is the same, e.g., both are adenine. In embodiments, the nucleiotide at the first and second position are different, e.g., one is adenine and one is thymine. In an embodiment, at least X% of the TREMs in a TREM composition has a non- naturally occurring modification at a first position and less than Y% have a non-naturally occurring modification at a second position, wherein X is 80, 90, 95, 96, 97, 98, 99, or 99.5 and Y is 20, 20, 5, 2, 1, 0.1, or 0.01.
  • the nucleotide at the first and second position is the same, e.g., both are adenine. In embodiments the nucleotide at the first and second position are different, e.g., one is adenine and one is thymine.
  • TREM, TREM core fragment and TREM fragment A “tRNA-based effector molecule” or “TREM” refers to an RNA molecule comprising one or more of the properties described herein.
  • the TREMs described herein comprise a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide.
  • a TREM comprising the LNA moiety or the 2’,5’-linked nucleotide can further comprise an additional non-naturally occurring modification, e.g., as provided in Table 5.
  • a TREM includes a TREM comprising a sequence of Formula A; a TREM core fragment comprising a sequence of Formula B; or a TREM fragment comprising a portion of a TREM which comprises a sequence of Formula A.
  • a TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]- [L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2].
  • [VL Domain] is optional.
  • a TREM fragment comprises a portion of a TREM, wherein the TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]- [ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2], and wherein the TREM fragment comprises: one, two, three or all or any combination of the following: a TREM half (e.g., from a cleavage in the ACH Domain, e.g., in the anticodon sequence, e.g., a 5’half or a 3’ half); a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DH Domain or the ACH Domain); a 3’ fragment (e.g., a fragment comprising the 3’ end, e.g., from a cleavage in
  • Exemplary TREM fragments include TREM halves (e.g., from a cleavage in the ACHD, e.g., 5’TREM halves or 3’ TREM halves), a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DHD or the ACHD), a 3’ fragment (e.g., a fragment comprising the 3’ end of a TREM, e.g., from a cleavage in the THD), or an internal fragment (e.g., from a cleavage in one or more of the ACHD, DHD or THD).
  • TREM halves e.g., from a cleavage in the ACHD, e.g., 5’TREM halves or 3’ TREM halves
  • a 5’ fragment e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DHD or
  • a TREM, a TREM core fragment or a TREM fragment can be charged with an amino acid (e.g., a cognate amino acid); charged with a non-cognate amino acid (e.g., a mischarged TREM (mTREM)); or not charged with an amino acid (e.g., an uncharged TREM (uTREM)).
  • an amino acid e.g., a cognate amino acid
  • mTREM mischarged TREM
  • uTREM uncharged TREM
  • a TREM, a TREM core fragment or a TREM fragment can be charged with an amino acid selected from alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.
  • a non-extended anticodon is an anticodon of no more than three nucleotides.
  • the TREM, TREM core fragment or TREM fragment is a cognate TREM. In an embodiment, the TREM, TREM core fragment or TREM fragment is a non- cognate TREM. In an embodiment, the TREM, TREM core fragment or TREM fragment recognizes a codon provided in Table 2 or Table 3.
  • Table 2 List of codons Attorney Docket No.: F2099-7037WO AAA AAC Attorney Docket No.: F2099-7037WO GGA GGC Table 3: Amino acids and corresponding codons Amino Acid mRNA codons Attorney Docket No.: F2099-7037WO Phenylalanine UUU, UUC Proline CCU, CCC, CCA, CCG , p leic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1- 451 disclosed in Table 1.
  • a TREM comprises an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM comprises an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM, a TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM, a TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM, a TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM core fragment or a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, Attorney Docket No.: F2099-7037WO 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM core fragment or a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM core fragment or a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM core fragment or a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM core fragment or a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM core fragment or a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence with at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identity to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1.
  • a TREM core fragment or a TREM fragment comprises a sequence of a length of between 10-90 ribonucleotides (rnt), between 10-80 rnt, between 10-70 rnt, between 10-60 rnt, between 10-50 rnt, between 10-40 rnt, between 10-30 rnt, between 10-20 rnt, between 20-90 rnt, between 20-80 rnt, 20-70 rnt, between 20-60 rnt, between 20-50 rnt, between 20-40 rnt, between 30-90 rnt, between 30-80 rnt, between 30-70 rnt, between 30-60 rnt, or between 30- 50 rnt.
  • rnt ribonucleotides
  • a TREM, a TREM core fragment or a TREM fragment described herein comprises a modification provided in Table 5, or a combination thereof.
  • the modifications provided in Table 5 are non-naturally occurring or occur naturally in RNAs, and are used herein on a synthetic TREM, a TREM core fragment or a TREM fragment at a position that does not occur in nature.
  • the TREM may comprise both LNA moieties and 2’,5’-linked nucleotides, optionally further comprising at least one additional non-naturally occurring modification
  • the TREM comprising the LNA moiety or the 2’,5’-linked nucleotide further comprises at least one additional non-naturally occurring modification (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications).
  • Locked Nucleic Acids Described herein are TREMs comprising a nucleotide comprising a locked nucleic acid moiety.
  • LNA locked nucleic acid
  • the LNA moiety refers to a nucleotide comprising a modified nucleotide sugar, in which the 2’-carbon is linked to the 4’-carbon atom through a cyclic structure.
  • the LNA moiety comprises a heterocyclyl ring or cycloalkyl ring between the 2’-position and the 4’-position of the nucleotide sugar within the Attorney Docket No.: F2099-7037WO nucleotide.
  • the TREM is a sequence of Formula (I), wherein the heterocyclyl ring comprises an oxygen atom, nitrogen atom, sulfur atom, selenium atom.
  • the heterocyclyl ring or cycloalkyl ring is a five-membered ring, six-membered ring, seven-membered ring, or eight-membered ring.
  • the present disclosure features a TREM containing a nucleotide comprising the LNA moiety having the structure of Formula (II): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a (R B ); each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, - OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substitute
  • B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil).
  • B is a purine-based nucleobase.
  • B is a pyrimidine-based nucleobase.
  • B is adenine.
  • B is cytosine.
  • B is guanine.
  • B is uracil.
  • B is a non-naturally occurring nucleobase.
  • B comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification).
  • the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. In some embodiments, the chemical modification is a nucleotide sugar modification. In some embodiments, the chemical modification is a 2’OMe modification. In some embodiments, the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification. In some embodiments, the chemical modification is a 2’MOE modification. In some embodiments, the chemical modification is a 2’-deoxy modification. In some embodiments, the chemical modification is present in the internucleotide region (e.g., a backbone modification).
  • the internucleotide region e.g., a backbone modification.
  • R 1 is C 1 -C 6 alkyl. In an embodiment, R 1 is C 1 - C6 heteroalkyl. In an embodiment, R 1 is C1-C6 haloalkyl. In an embodiment, R 1 is halo. I n some embodiments, R2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an Attorney Docket No.: F2099-7037WO embodiment, R 2 is hydrogen. In an embodiment, R 2 is C 1 -C 6 alkyl.
  • R 2 is C 1 - C6 heteroalkyl. In an embodiment, R 2 is C1-C6 haloalkyl. In an embodiment, R 2 is halo. In some embodiments, R 3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, R 3 is hydrogen. In an embodiment, R 3 is C1-C6 alkyl. In an embodiment, R 3 is C1- C6 heteroalkyl. In an embodiment, R 3 is C1-C6 haloalkyl.
  • R 3 is halo.
  • each of R 4a and R 4b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • each of R 4a and R 4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl.
  • each of R 4a and R 4b is independently hydrogen.
  • each of R 4a and R 4b is independently C 1 -C 6 alkyl.
  • each of R 4a and R 4b is independently C 1 -C 6 heteroalkyl.
  • one of L, M, Z, and X is N(R B )-. In some embodiments, one of L and X is O. In some embodiments, one of L and X is N(R B )-. In some embodiments, X is selected from C1-C6 alkylene, O, N(R B ), S(R D )x-, and -Se(R E ) y -. In some embodiments, X is selected from C 1 -C 6 alkylene, O, N(R B ), S, and Se. In some embodiments, X is O. In some embodiments, X is C 1 -C 6 alkylene. In some embodiments, X is N(R B ).
  • X is S. In some embodiments, X is Se.
  • the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-a): Y Attorney Docket No.: F2099-7037WO is C(R’), O, or N(R B ); each of R 1 , R 2 , and R 3 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C 1 -C 6 alkylene, C 2 -C 6 alkenylene,
  • Y is selected from CH2, S, or O. In an embodiment, Y is CH2. In an embodiment, Y is S. In an embodiment, Y is O.
  • R 1 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R1 is hydrogen. In an embodiment, R1 is C 1 -C 6 alkyl. In an embodiment, R1 is C 1 - C 6 heteroalkyl. In an embodiment, R 1 is C 1 -C 6 haloalkyl.
  • R 3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R 3 is hydrogen. In an embodiment, R 3 is C 1 -C 6 alkyl. In an embodiment, R 3 is C 1 - C6 heteroalkyl. In an embodiment, R 3 is C1-C6 haloalkyl. In an embodiment, R 3 is halo.
  • each of R 4a and R 4b is independently halo. In an embodiment, one of R 4a and R 4b is independently hydrogen. In an embodiment, one of R 4a and R 4b is independently C1-C6 alkyl. In an embodiment, one of R 4a and R 4b is independently C 1 -C 6 heteroalkyl. In an embodiment, one of R 4a and R 4b is independently C 1 -C 6 haloalkyl. In an embodiment, one of R 4a and R 4b is independently halo. In some embodiments, one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(R B )-. In some embodiments, one of L and X is O.
  • one of L and X is N(R B )-.
  • X is selected from C 1 -C 6 alkylene, O, N(R B ), S(R D ) x -, and -Se(R E )y-.
  • X is selected from C1-C6 alkylene, O, N(R B ), S, and Se.
  • X is O.
  • X is C1-C6 alkylene.
  • X is N(R B ).
  • X is S.
  • X is Se.
  • each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, each of R ia and R ib is independently hydrogen. In an embodiment, each of R ia and R ib is independently C 1 -C 6 alkyl. In an embodiment, each of R ia and R ib is independently C1-C6 heteroalkyl.
  • each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R ii and R iii is independently hydrogen. In an embodiment, each of R ii and R iii is independently C1-C6 alkyl. In an embodiment, each of R ii and R iii is independently C 1 -C 6 heteroalkyl.
  • each of R ii and R iii is independently C1-C6 haloalkyl. In an embodiment, each of R ii and R iii is independently halo. In an embodiment, one of R ii and R iii is independently hydrogen. In an embodiment, one of R ii and R iii is independently C 1 -C 6 alkyl. In an embodiment, one of R ii and R iii is independently C 1 -C 6 heteroalkyl. In an embodiment, one of R ii and R iii is independently C1-C6 haloalkyl. In an embodiment, one of R ii and R iii is independently halo.
  • the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-b): a salt, stereoisomer, tautomer, or hydrate thereof
  • R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, -OR A , or - N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N
  • R 1 is halo.
  • R 2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • R 2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl.
  • R 2 is hydrogen.
  • R 2 is C 1 -C 6 alkyl.
  • R 2 is C 1 - C 6 heteroalkyl.
  • R 2 is C 1 -C 6 haloalkyl.
  • R 2 is halo.
  • R 3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, R 3 is hydrogen. In an embodiment, R 3 is C 1 -C 6 alkyl. In an embodiment, R 3 is C 1 - C6 heteroalkyl. In an embodiment, R 3 is C1-C6 haloalkyl. In an embodiment, R 3 is halo.
  • one of L, M, Z, and X is N(R B )-. In some embodiments, one of L and X is O. In some embodiments, one of L and X is N(R B )-. In some embodiments, X is selected from C 1 -C 6 alkylene, O, N(R B ), S(R D ) x -, and -Se(R E )y-. In some embodiments, X is selected from C1-C6 alkylene, O, N(R B ), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(R B ).
  • X is S. In some embodiments, X is Se.
  • each of R ia and R ib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl.
  • each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl.
  • each of R ia and R ib is independently hydrogen. In an embodiment, each of R ia and R ib is independently C 1 -C 6 alkyl. In an embodiment, each of R ia and R ib is independently C1-C6 heteroalkyl.
  • each of R ia and R ib is independently C 1 -C 6 haloalkyl. In an embodiment, one of R ia and R ib is independently hydrogen. In an embodiment, one of R ia and R ib is independently C 1 -C 6 alkyl. In an embodiment, one of R ia and R ib is independently C1-C6 heteroalkyl. In an embodiment, one of R ia and R ib is independently C1-C6 haloalkyl. In some embodiments, each of R ii and R iii is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R ii and R iii is independently hydrogen. In an embodiment, each of R ii and R iii is independently C 1 -C 6 alkyl. In an embodiment, each of R ii and R iii is independently C 1 -C 6 heteroalkyl. In an embodiment, each of R ii and R iii is independently C1-C6 haloalkyl. In an embodiment, each of R ii and R iii is independently halo. In an embodiment, one of Rii and Riii is independently hydrogen.
  • one of R ii and R iii is independently C 1 -C 6 alkyl. In an embodiment, one of R ii and Attorney Docket No.: F2099-7037WO R iii is independently C 1 -C 6 heteroalkyl. In an embodiment, one of R ii and R iii is independently C1-C6 haloalkyl. In an embodiment, one of R ii and R iii is independently halo.
  • the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-c): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y is and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C 1 -C 6 alkylene, C 2 -C 6 alkenylene, C 2 -C 6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -
  • Y is selected from CH 2 , S, or O. In an embodiment, Y is CH 2 . In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R 1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, R 1 is hydrogen. In an embodiment, R 1 is C 1 -C 6 alkyl. In an embodiment, R 1 is C 1 - C6 heteroalkyl. In an embodiment, R 1 is C1-C6 haloalkyl.
  • R 1 is halo.
  • R 2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl.
  • R 2 is hydrogen.
  • R 2 is C1-C6 alkyl.
  • R 2 is C1- C6 heteroalkyl.
  • R 2 is C1-C6 haloalkyl.
  • R 2 is halo.
  • R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, R 3 is hydrogen. In an embodiment, R 3 is C1-C6 alkyl. In an embodiment, R 3 is C1- C 6 heteroalkyl. In an embodiment, R 3 is C 1 -C 6 haloalkyl. In an embodiment, R 3 is halo.
  • each of R 4a and R 4b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R 4a and R 4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R 4a and R 4b is independently hydrogen. In an embodiment, each of R 4a and R 4b is independently C 1 -C 6 alkyl. In an embodiment, each of R 4a and R 4b is independently C 1 -C 6 heteroalkyl. In an embodiment, each of R 4a and R 4b is independently C1-C6 haloalkyl.
  • each of R 4a and R 4b is independently halo. In an embodiment, one of R 4a and R 4b is independently hydrogen. In an embodiment, one of R 4a and R 4b is independently C 1 -C 6 alkyl. In an embodiment, one of R 4a and R 4b is independently C1-C6 heteroalkyl. In an embodiment, one of R 4a and R 4b is independently C 1 -C 6 haloalkyl. In an embodiment, one of R4a and R4b is independently halo.
  • one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(R B )-.
  • one of L and X is O. In some embodiments, one of L and X is N(R B )-. In some embodiments, X is selected from C1-C6 alkylene, O, N(R B ), S(R D )x-, and -Se(R E ) y -. In some embodiments, X is selected from C 1 -C 6 alkylene, O, N(R B ), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(R B ). In some embodiments, X is S. In some embodiments, X is Se.
  • each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, each of R ia and R ib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R ia and R ib is independently hydrogen. In an embodiment, each of R ia and R ib is independently C 1 -C 6 alkyl. In an embodiment, each of R ia and R ib is independently C 1 -C 6 heteroalkyl.
  • each of R ia and R ib is independently C1-C6 haloalkyl. In an embodiment, one of R ia and R ib is independently hydrogen. In an embodiment, one of R ia and R ib is independently C1-C6 alkyl. In an embodiment, one of R ia and R ib is independently C 1 -C 6 heteroalkyl. In an embodiment, one of R ia and R ib is independently C1-C6 haloalkyl. In some embodiments, each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • each of R ii and R iii is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, each of R ii and R iii is independently hydrogen. In an embodiment, each of R ii and R iii is independently C1-C6 alkyl. In an embodiment, each of R ii and R iii is independently C 1 -C 6 heteroalkyl. In an embodiment, each of R ii and R iii is independently C 1 -C 6 haloalkyl. In an embodiment, each of R ii and R iii is independently halo. In an embodiment, one of R ii and R iii is independently hydrogen.
  • one of R ii and R iii is independently C1-C6 alkyl. In an embodiment, one of R ii and R iii is independently C 1 -C 6 heteroalkyl. In an embodiment, one of R ii and R iii is independently C 1 -C 6 haloalkyl. In an embodiment, one of R ii and R iii is independently halo.
  • the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-d): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y R 2 , and R 3 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 C 1 -C 6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C 1 -C 6 heteroalkylene C 1 -C 6 haloalkylene -O-, -N(R B )-
  • R 1 is halo.
  • R 2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl.
  • R 2 is hydrogen.
  • R 2 is C 1 -C 6 alkyl.
  • R 2 is C 1 - C6 heteroalkyl.
  • R 2 is C1-C6 haloalkyl.
  • R 2 is halo.
  • each of R 4a and R 4b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R 4a and R 4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R 4a and R 4b is independently hydrogen. In an embodiment, each of R 4a and R 4b is independently C 1 -C 6 alkyl. In an embodiment, each of R 4a and R 4b is independently C 1 -C 6 heteroalkyl. In an embodiment, each of R 4a and R 4b is independently C1-C6 haloalkyl.
  • each of R 4a and R 4b is independently halo. In an embodiment, one of R 4a and R 4b is independently hydrogen. In an embodiment, one of R 4a and R 4b is independently C 1 -C 6 alkyl. In an embodiment, one of R 4a and R 4b is independently C 1 -C 6 heteroalkyl. In an embodiment, one of R 4a and R 4b is independently C1-C6 haloalkyl. In an embodiment, one of R 4a and R 4b is independently halo. In some embodiments, one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(R B )-. In some embodiments, one of L and X is O.
  • one of L and X is N(R B )-.
  • X is selected from C1-C6 alkylene, O, N(R B ), S(R D )x-, and -Se(RE) y -.
  • X is selected from C 1 -C 6 alkylene, O, N(RB ), S, and Se.
  • X is O.
  • X is C 1 -C 6 alkylene.
  • X is N(R B ).
  • X is S.
  • X is Se.
  • R i is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R i is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, R i is hydrogen. In an embodiment, R i is C1-C6 alkyl. In an embodiment, R i is C1-C6 heteroalkyl. In an embodiment, R i is C1-C6 haloalkyl.
  • each of R ii and R iii is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R ii and R iii is independently hydrogen. In an embodiment, each of R ii and R iii is independently C 1 -C 6 alkyl. In an embodiment, each of R ii and R iii is independently C 1 -C 6 heteroalkyl.
  • each of R ii and R iii is independently C1-C6 haloalkyl. In an embodiment, each of R ii and R iii is independently halo. In an embodiment, one of R ii and R iii is independently hydrogen. In an embodiment, one of R ii and R iii is independently C 1 -C 6 alkyl. In an embodiment, one of R ii and R iii is independently C1-C6 heteroalkyl. In an embodiment, one of R ii and R iii is independently C1-C6 haloalkyl. In an embodiment, one of R ii and R iii is independently halo.
  • the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-e): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a ; each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, - OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; X is C 1 -C 6 alkylene, C 2 -C 6 alkenylene, C 2 -C 6 alkynylene, C1-C6 heteroalkylene C1-C6 haloal
  • Y is selected from CH 2 , S, or O. In an embodiment, Y is CH 2 . In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R 1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, R 1 is hydrogen. In an embodiment, R 1 is C 1 -C 6 alkyl. In an embodiment, R 1 is C 1 - C6 heteroalkyl. In an embodiment, R 1 is C1-C6 haloalkyl.
  • R 1 is halo.
  • R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl.
  • R 2 is hydrogen.
  • R 2 is C1-C6 alkyl.
  • R 2 is C1- C6 heteroalkyl.
  • R 2 is C1-C6 haloalkyl.
  • R 2 is halo.
  • R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, R 3 is hydrogen. In an embodiment, R 3 is C1-C6 alkyl. In an embodiment, R 3 is C1- C 6 heteroalkyl. In an embodiment, R 3 is C 1 -C 6 haloalkyl. In an embodiment, R 3 is halo.
  • each of R 4a and R 4b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R 4a and R 4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R 4a and R 4b is independently hydrogen. In an embodiment, each of R 4a and R 4b is independently C 1 -C 6 alkyl. Attorney Docket No.: F2099-7037WO In an embodiment, each of R 4a and R 4b is independently C 1 -C 6 heteroalkyl.
  • each of R 4a and R 4b is independently C1-C6 haloalkyl. In an embodiment, each of R 4a and R 4b is independently halo. In an embodiment, one of R 4a and R 4b is independently hydrogen. In an embodiment, one of R 4a and R 4b is independently C 1 -C 6 alkyl. In an embodiment, one of R 4a and R 4b is independently C1-C6 heteroalkyl. In an embodiment, one of R 4a and R 4b is independently C1-C6 haloalkyl. In an embodiment, one of R 5a and R 5b is independently halo.
  • each of R 5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R 5a and R 5b is independently hydrogen. In an embodiment, each of R 5a and R 5b is independently C 1 -C 6 alkyl. In an embodiment, each of R 5a and R 5b is independently C 1 -C 6 heteroalkyl. In an embodiment, each of R 5a and R 5b is independently C1-C6 haloalkyl.
  • each of R 5a and R 5b is independently halo. In an embodiment, one of R 5a and R 5b is independently hydrogen. In an embodiment, one of R 5a and R 5b is independently C 1 -C 6 alkyl. In an embodiment, one of R 5a and R 5b is independently C1-C6 heteroalkyl. In an embodiment, one of R 5a and R 5b is independently C1-C6 haloalkyl. In an embodiment, one of R 5a and R 5b is independently halo. In some embodiments, X is selected from C 1 -C 6 alkylene, O, N(R B ), S(R D ) x -, and - Se(R E ) y -.
  • X is selected from C 1 -C 6 alkylene, O, N(R B ), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(R B ). In some embodiments, X is S. In some embodiments, X is Se. In some embodiments, n is selected from 1, 2, or 3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6.
  • the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-e-i) or (II-e-ii): e-ii), or a salt, stereoisomer, Attorney Docket No.: F2099-7037WO
  • the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-f): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; X is C1-C6 al
  • the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-f-i) or (II-f-ii): Attorney Docket No.: F2099-7037WO f-ii), or a salt, stereoisomer, some sequencing comprising the LNA moiety having a structure of Formula (II-f) is selected from: or
  • the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-g): a salt, stereoisomer, tautomer, or hydrate thereof, B is a nucleobase; each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R
  • the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-g-i) or Formula (II-g-ii): g-ii), or a salt, stereoisomer, tautomer, or In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-g-iii): iii), or a salt, stereoisomer, tautomer, or hydrate thereof.
  • each of R 1 , R 2 , and R 3 is independently C 6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ;
  • X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C 1 -C 6 heteroalkylene C 1 -C 6 haloalkylene -O-, -N(R B )-
  • the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-m): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof,
  • R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ;
  • each of R 4a and R 4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -
  • the nucleotide within the TREM sequence comprising the LNA moiety having a structure of Formula (II-l) is selected from: Attorney Docket No.: F2099-7037WO moiety has a structure of Formula (II-m-i): a salt, stereoisomer, tautomer, or hydrate thereof.
  • each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ;
  • X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N
  • each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R 5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl. In an embodiment, each of R 5a and R 5b is independently hydrogen. In an embodiment, each of R 5a and R 5b is independently C 1 -C 6 alkyl. In an embodiment, each of R 5a and R 5b is independently C1-C6 heteroalkyl. In an embodiment, each of R 5a and R 5b is independently C 1 -C 6 haloalkyl.
  • each of R 5a and R 5b is independently halo. In an embodiment, one of R 5a and R 5b is independently hydrogen. In an embodiment, one of R 5a and R 5b is independently C1-C6 alkyl. In an embodiment, one of R 5a and R 5b is independently C1-C6 heteroalkyl. In an embodiment, one of R 5a and R 5b is independently C 1 -C 6 haloalkyl. In an embodiment, one of R 5a and R 5b is independently halo.
  • each of R 6a and R 6b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R 6a and R 6b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R 6a and R 6b is independently hydrogen. In an embodiment, each of R 6a and R 6b is independently C 1 -C 6 alkyl. In an embodiment, each of R 6a and R 6b is independently C1-C6 heteroalkyl.
  • each of R 6a and R 6b is independently C1-C6 haloalkyl. In an embodiment, each of R 6a and R 6b is independently halo. In an embodiment, one of R 6a and R 6b is independently hydrogen. In an embodiment, one of R 6a and R 6b is independently C1-C6 alkyl. In an embodiment, one of R 6a and R 6b is independently C1-C6 heteroalkyl. In an embodiment, one of R 6a and R 6b is independently C 1 -C 6 haloalkyl. In an embodiment, one of R 6a and R 6b is independently halo.
  • the nucleotide within the TREM sequence comprising the LNA moiety has the structure of Formula (II-s) or Formula (II-t): a salt, stereoisomer, tautomer,
  • the TREM comprises a single LNA moiety.
  • the TREM comprises a plurality of LNA moieties.
  • the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 LNA moieties.
  • the TREM comprises 2 LNA moieties.
  • the TREM comprises 3 LNA moieties.
  • the TREM comprises 4 LNA moieties.
  • the TREM comprises 5 LNA moieties. In some embodiments, the TREM comprises 6 LNA moieties. In some embodiments, the TREM comprises at least 7 LNA moieties. In some embodiments, the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. In some embodiments, the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. In some embodiments, the LNA moiety is present in the [L2]-[DH Domain]-[L3]. In some embodiments, the LNA moiety is present in the [L3]-[ACH Domain].
  • the LNA moiety Attorney Docket No.: F2099-7037WO is present in the [VL Domain]. In some embodiments, the LNA moiety is present in the [TH Domain]. In some embodiments, the LNA moiety is present in the [L4]-[ASt Domain2]-[L5]. In some embodiments, the TREM comprises a locked nucleic acid in the [ASt Domain1]. In some embodiments, the TREM comprises a locked nucleic acid in the [DH Domain]. In some embodiments, the TREM comprises a locked nucleic acid in the [ACH Domain]. In some embodiments, the TREM comprises a locked nucleic acid in the [VL Domain].
  • the TREM comprises a locked nucleic acid in the [TH Domain]. In some embodiments, the TREM comprises a locked nucleic acid in the [ASt Domain2]. In an embodiment, the LNA moiety is present at any one of nucleotide positions 1, 14, 15, 16, 28, 29, 43, 44, 45, 54, 65, 68, and 70 of the TREM. In an embodiment, the locked nucleic acid is present at any one of nucleotide position 14, 15, 44, or 54 of the TREM. In an embodiment, the locked nucleic acid is not present at one of the nucleotide positions 41, 42, or 63 of the TREM. In an embodiment, the locked nucleic acid is not present at the nucleotide position 26 of the TREM.
  • the TREM comprises two or more locked nucleic acids. In some embodiments, the TREM comprises two locked nucleic acids. In some embodiments, the TREM comprises two locked nucleic acids, each independently located in the [ASt Domain1] and the [VL Domain]. In some embodiments, the TREM comprises two locked nucleic acids, each independently located in the [ASt Domain1] and the [TH Domain]. In some embodiments, the TREM comprises two locked nucleic acids, each independently located in the [ASt Domain1] and the [ASt Domain2].
  • the TREM comprises two locked nucleic acids, each independently located at one of nucleotide positions 14, 15, 44, 54, and 65 (e.g., 14 and 15, e.g., 15 and 65, e.g., 14 and 65, or e.g., 14 and 54).
  • the TREM comprises three locked nucleic acids.
  • the TREM comprises three locked nucleic acids each independently located in the [ACH Domain], the [TH Domain], and the [ASt Domain2].
  • the TREM comprises one locked nucleic acid located in the [ACH Domain], and two locked nucleic acids in the [VL Domain].
  • the TREM comprises one locked nucleic acid in the [TH Domain], and two locked nucleic acids in the [DH Domain]. In some embodiments, the TREM comprises three locked nucleic acids, each independently located at any of the nucleotide positions 14, 15, 54, or 65 (e.g., 14, 15, and 54, e.g., 14, 15, and 65). In some embodiments, the Attorney Docket No.: F2099-7037WO TREM comprises three locked nucleic acids, each independently located at nucleotide position 14, 15, and 54. In some embodiments, the TREM comprises three locked nucleic acids, each independently located at nucleotide position 14, 15, and 65.
  • the TREM comprises four locked nucleic acids located in the [DH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM comprises four locked nucleic acids located in the [DH Domain], the [ACH Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM comprises four locked nucleic acids, each independently located at nucleotide position 14, 44, 54, and 65. In some embodiments, the TREM comprises four locked nucleic acids, each independently located at nucleotide position 14, 43, 54, and 65. In some embodiments, the TREM comprises five locked nucleic acids. In some embodiments, the TREM comprises six locked nucleic acids.
  • the TREM comprises seven or more locked nucleic acids.
  • the TREM is a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 625-793, 992-1252, 1357, 1364, 1378-1393, or 1397-1398 (e.g., any one of SEQ ID Nos: 625-793, e.g., any one of SEQ ID Nos: 992-1252, 1357, 1364, 1378-1393, or 1397-1398).
  • the TREM is a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793, 992-1252, 1357, 1364, 1378- 1393, and 1397-1398 (e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793, or a TREM having a sequence of any one of SEQ ID NOs: 992-1252, 1357, 1364, 1378-1393, and 1397-1398).
  • a TREM having a sequence of any one of SEQ ID NOs: 626-793, 992-1252, 1357, 1364, 1378-1393, and 1397-1398 e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793, or a TREM having a sequence of any one of SEQ ID NOs: 992-1252, 1357, 1364, 1378-1393, and 1397-1398.
  • the TREM is a TREM having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or more sequence identity to a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793.
  • the TREM is a TREM having at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 1617, 18, 19, 20, or more nucleotides that differ from a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793.
  • the TREM is a TREM having 1-5, 5-10, 10-15, or 15-20 nucleotides that differ from a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793.
  • the TREM is a TREM having at least one additional non-naturally occurring modification compared with a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793.
  • the TREM is a TREM having at least 2, 3, 4, 5, 6, 7, 8.9, 10, or Attorney Docket No.: F2099-7037WO more additional non-naturally occurring modifications compared with a TREM provided in FIG. 1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793.
  • the locked nucleic acid is located at the nucleotide position 14, 15, 44, 54, or 65 of the TREM. In an embodiment, the locked nucleic acid is located at the nucleotide position 14 of the TREM. In an embodiment, the locked nucleic acid is located at nucleotide position 15 of the TREM.
  • the locked nucleic acid is located at the nucleotide position 54 of the TREM. In an embodiment, the locked nucleic acid is located at the nucleotide position 65 of the TREM. In an embodiment, the locked nucleic acid is not located at the nucleotide position 41, 42, or 63 of the TREM. In an embodiment, the locked nucleic acid is not located at the nucleotide position 7, 8, 9, 10, 17, 18, 31, 32, 33, 34, 35, 36, 37, 38, 39, 48, 55, 58, 59, 60, 61, 72, or 73 (e.g., 7-10, 17-18, 31-39, 48, 55, 58-61, or 72-73).
  • the locked nucleic acid is not located at the nucleotide positions 7-10 (e.g., 7, 8, 9, or 10). In an embodiment, the locked nucleic acid is not located at the nucleotide position 7. In an embodiment, the locked nucleic acid is not located at the nucleotide position 8. In an embodiment, the locked nucleic acid is not located at the nucleotide position 9. In an embodiment, the locked nucleic acid is not located at the nucleotide position 10. In an embodiment, the locked nucleic acid is not located at the nucleotide position 17-18 (e.g., 17 or 18). In an embodiment, the locked nucleic acid is not located at the nucleotide position 17.
  • the locked nucleic acid is not located at the nucleotide position 18. In an embodiment, the locked nucleic acid is not located at the nucleotide position 31-39 (e.g., 31, 32, 33, 34, 35, 36, 37, 38, or 39). In an embodiment, the locked nucleic acid is not located at the nucleotide position 31. In an embodiment, the locked nucleic acid is not located at the nucleotide position 32. In an embodiment, the locked nucleic acid is not located at the nucleotide position 33. In an embodiment, the locked nucleic acid is not located at the nucleotide position 34. In an embodiment, the locked nucleic acid is not located at the nucleotide position 35.
  • the locked nucleic acid is not located at the nucleotide position 31-39 (e.g., 31, 32, 33, 34, 35, 36, 37, 38, or 39). In an embodiment, the locked nucleic acid is not located at the nucleotide position 31. In an embodiment,
  • the locked nucleic acid is not located at the nucleotide position 36. In an embodiment, the locked nucleic acid is not located at the nucleotide position 37. In an embodiment, the locked nucleic acid is not located at the nucleotide position 38. In an embodiment, the locked nucleic acid is not located at the nucleotide position 39.
  • Attorney Docket No.: F2099-7037WO In an embodiment, the locked nucleic acid is not located at the nucleotide position 48. In an embodiment, the locked nucleic acid is not located at the nucleotide position 55.
  • the locked nucleic acid is not located at the nucleotide position 58-61 (e.g., 58, 59, 60, or 61). In an embodiment, the locked nucleic acid is not located at the nucleotide position 58. In an embodiment, the locked nucleic acid is not located at the nucleotide position 59. In an embodiment, the locked nucleic acid is not located at the nucleotide position 60. In an embodiment, the locked nucleic acid is not located at the nucleotide position 61. In an embodiment, the locked nucleic acid is not located at the nucleotide position 72-73 (e.g., 72 or 73).
  • the locked nucleic acid is not located at the nucleotide position 72. In an embodiment, the locked nucleic acid is not located at the nucleotide position 73.
  • the TREM comprises the non-naturally occurring modification pattern of TREM No.166 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 12-m, 13-f, 14-LNA, 16-m, 17-m, 19-*, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 41-m, 42- m, 43-LNA, 44-f, 45-LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 60- f, 61-m, 62-m, 63-f*, 64-d, 65
  • the TREM comprises the non-naturally occurring modification pattern of TREM No.167 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 12-m, 13-f, 14-LNA, 15-f, 16-m, 17-m, 19-*, 21-f, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 40-f, 41-m, 42-m, 43-LNA, 44-f, 45-LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 59-f, 60-f, 61-m, 62-m, 63-f*, 64-d, 65-LNA, 66-m, 67-m, 68-f, 71-m, 72-f, 73-m
  • the TREM comprises the non-naturally occurring modification pattern of TREM No.168 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 13-f, 15-f, 17- m, 19-*, 20-d, 21-f, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 40-f, 41-m, 42-m, 43-LNA, 44-f, 45-LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 59-f, 60-f, 61-m, 62-m, 63-f*, 64-d, 65-LNA, 66-m, 67-m, 68-f, 72-f, 73-m, 75-f*.
  • the TREM comprises the non-naturally occurring modification pattern of TREM No.169 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 13-f, 17-m, 19-*, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 41-m, 42-m, 43-LNA, 44-f, 45- Attorney Docket No.: F2099-7037WO LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 60-f, 61-m, 62-m, 63-f*, 64-d, 65-LNA, 66-m, 67-m, 68-f, 72-f, 73-m, 75-f*.
  • F2099-7037WO LNA 46-d, 49-m
  • the TREM containing an LNA moiety is TREM No.15. In some embodiments, the TREM containing an LNA moiety is TREM No.16. In some embodiments, the TREM containing an LNA moiety is TREM No.43. In some embodiments, the TREM containing an LNA moiety is TREM No.44. In some embodiments, the TREM containing an LNA moiety is TREM No.45. In some embodiments, the TREM containing an LNA moiety is TREM No.46. In some embodiments, the TREM containing an LNA moiety is TREM No.66. In some embodiments, the TREM containing an LNA moiety is TREM No.79.
  • the TREM containing an LNA moiety is TREM No.80. In some embodiments, the TREM containing an LNA moiety is TREM No.81. In some embodiments, the TREM containing an LNA moiety is TREM No.82. In some embodiments, the TREM containing an LNA moiety is TREM No.83. In some embodiments, the TREM containing an LNA moiety is TREM No.84. In some embodiments, the TREM containing an LNA moiety is TREM No.85. In some embodiments, the TREM containing an LNA moiety is TREM No.86. In some embodiments, the TREM containing an LNA moiety is TREM No.87.
  • the TREM containing an LNA moiety is TREM No.88. In some embodiments, the TREM containing an LNA moiety is TREM No.89. In some embodiments, the TREM containing an LNA moiety is TREM No.90. In some embodiments, the TREM containing an LNA moiety is TREM No.91. In some embodiments, the TREM containing an LNA moiety is TREM No.92. In some embodiments, the TREM containing an LNA moiety is TREM No.93. In some embodiments, the TREM containing an LNA moiety is TREM No.94. In some embodiments, the TREM containing an LNA moiety is TREM No.95.
  • the TREM containing an LNA moiety is TREM No.96. In some embodiments, the TREM containing an LNA moiety is TREM No.97. In some embodiments, the TREM containing an LNA moiety is TREM No.98. In some embodiments, the TREM containing an LNA moiety is TREM No.99. In some embodiments, the TREM containing an LNA moiety is TREM No.100. In some embodiments, the TREM containing an LNA moiety is TREM No.101. In some embodiments, the TREM containing an LNA moiety is TREM No.102. In some embodiments, the TREM containing an LNA moiety is TREM No.103.
  • the TREM containing an LNA moiety is TREM No.104. In some embodiments, the TREM containing an LNA moiety is Attorney Docket No.: F2099-7037WO TREM No.105. In some embodiments, the TREM containing an LNA moiety is TREM No.106. In some embodiments, the TREM containing an LNA moiety is TREM No.107. In some embodiments, the TREM containing an LNA moiety is TREM No.108. In some embodiments, the TREM containing an LNA moiety is TREM No.109. In some embodiments, the TREM containing an LNA moiety is TREM No.110. In some embodiments, the TREM containing an LNA moiety is TREM No.111.
  • the TREM containing an LNA moiety is TREM No.112. In some embodiments, the TREM containing an LNA moiety is TREM No.113. In some embodiments, the TREM containing an LNA moiety is TREM No.114. In some embodiments, the TREM containing an LNA moiety is TREM No.115. In some embodiments, the TREM containing an LNA moiety is TREM No.116. In some embodiments, the TREM containing an LNA moiety is TREM No.117. In some embodiments, the TREM containing an LNA moiety is TREM No.118. In some embodiments, the TREM containing an LNA moiety is TREM No.120.
  • the TREM containing an LNA moiety is TREM No.121. In some embodiments, the TREM containing an LNA moiety is TREM No.122. In some embodiments, the TREM containing an LNA moiety is TREM No.123. In some embodiments, the TREM containing an LNA moiety is TREM No.124. In some embodiments, the TREM containing an LNA moiety is TREM No.125. In some embodiments, the TREM containing an LNA moiety is TREM No.126. In some embodiments, the TREM containing an LNA moiety is TREM No.127. In some embodiments, the TREM containing an LNA moiety is TREM No.128.
  • the TREM containing an LNA moiety is TREM No.129. In some embodiments, the TREM containing an LNA moiety is TREM No.130. In some embodiments, the TREM containing an LNA moiety is TREM No.132. In some embodiments, the TREM containing an LNA moiety is TREM No.133. In some embodiments, the TREM containing an LNA moiety is TREM No.134. In some embodiments, the TREM containing an LNA moiety is TREM No.135. In some embodiments, the TREM containing an LNA moiety is TREM No.136. In some embodiments, the TREM containing an LNA moiety is TREM No.137.
  • the TREM containing an LNA moiety is TREM No.138. In some embodiments, the TREM containing an LNA moiety is TREM No.139. In some embodiments, the TREM containing an LNA moiety is TREM No.140. In some embodiments, the TREM containing an LNA moiety is TREM No.141. In some embodiments, the TREM containing an LNA moiety is TREM No.142. In some embodiments, the TREM containing an LNA moiety is TREM No.143. Attorney Docket No.: F2099-7037WO In some embodiments, the TREM containing an LNA moiety is TREM No.144. In some embodiments, the TREM containing an LNA moiety is TREM No.145.
  • the TREM containing an LNA moiety is TREM No.146. In some embodiments, the TREM containing an LNA moiety is TREM No.147. In some embodiments, the TREM containing an LNA moiety is TREM No.148. In some embodiments, the TREM containing an LNA moiety is TREM No.149. In some embodiments, the TREM containing an LNA moiety is TREM No.150. In some embodiments, the TREM containing an LNA moiety is TREM No.151. In some embodiments, the TREM containing an LNA moiety is TREM No.152. In some embodiments, the TREM containing an LNA moiety is TREM No.153.
  • the TREM containing an LNA moiety is TREM No.154. In some embodiments, the TREM containing an LNA moiety is TREM No.155. In some embodiments, the TREM containing an LNA moiety is TREM No.156. In some embodiments, the TREM containing an LNA moiety is TREM No.157. In some embodiments, the TREM containing an LNA moiety is TREM No.158. In some embodiments, the TREM containing an LNA moiety is TREM No.159. In some embodiments, the TREM containing an LNA moiety is TREM No.160. In some embodiments, the TREM containing an LNA moiety is TREM No.161.
  • the TREM containing an LNA moiety is TREM No.162. In some embodiments, the TREM containing an LNA moiety is TREM No.163. In some embodiments, the TREM containing an LNA moiety is TREM No.164. In some embodiments, the TREM containing an LNA moiety is TREM No.165. In some embodiments, the TREM containing an LNA moiety is TREM No.166. In some embodiments, the TREM containing an LNA moiety is TREM No.444. In some embodiments, the TREM containing an LNA moiety is TREM No.445. In some embodiments, the TREM containing an LNA moiety is TREM No.451.
  • the TREM containing an LNA moiety is TREM No.454. In some embodiments, the TREM containing an LNA moiety is TREM No.547. In some embodiments, the TREM containing an LNA moiety is TREM No.566. In some embodiments, the TREM containing an LNA moiety is TREM No.628.
  • the present disclosure consists of a TREM sequence comprising the 2’,5’- linked nucleotide having a structure of Formula (III): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a ; each of R 1 , R 2 , and R 4 is independently hydrogen, C1-C6 C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or - N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, C2- C6 alkenyl, C2-C6 alkynyl, C1-C6
  • B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil).
  • B is a purine-based nucleobase.
  • B is a pyrimidine-based nucleobase.
  • B is adenine.
  • B is cytosine.
  • B is guanine.
  • B is uracil.
  • B is a non-naturally occurring nucleobase.
  • B comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification).
  • the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification.
  • the chemical Attorney Docket No.: F2099-7037WO modification is a nucleotide sugar modification.
  • the chemical modification is a 2’OMe modification.
  • the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification.
  • the chemical modification is a 2’MOE modification.
  • the chemical modification is a 2’-deoxy modification.
  • the chemical modification is present in the internucleotide region (e.g., a backbone modification).
  • the chemical modification is a phosphorothioate modification.
  • B does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification).
  • Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(R B ).
  • R 1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R 1 is hydrogen.
  • R 1 is C 1 -C 6 alkyl. In some embodiments, R 1 is C2-C6 alkenyl. In some embodiments, R 1 is C2-C6 alkynyl. In some embodiments, R 1 is C1- C6 heteroalkyl. In some embodiments, R 1 is C1-C6 haloalkyl. In some embodiments, R 1 is halo. In some embodiments, R 1 is cyano. In some embodiments, R 1 is -OR A . In some embodiments, R 1 is -N(R B )(R C ).
  • R 2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is C 1 -C 6 alkyl. In some embodiments, R 2 is C2-C6 alkenyl. In some embodiments, R 2 is C2-C6 alkynyl. In some embodiments, R 2 is C1- C6 heteroalkyl. In some embodiments, R 2 is C1-C6 haloalkyl. In some embodiments, R 2 is halo.
  • R 2 is cyano. In some embodiments, R 2 is -OR A . In some embodiments, R 2 is -N(R B )(R C ). In some embodiments, R 4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 4 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is C 1 -C 6 alkyl. In some embodiments, R 4 is C2-C6 alkenyl. In some embodiments, R 4 is C2-C6 alkynyl.
  • R 4 is C1- C 6 heteroalkyl. In some embodiments, R4 is C 1 -C 6 haloalkyl. In some embodiments, R4 is halo. Attorney Docket No.: F2099-7037WO In some embodiments, R 4 is cyano. In some embodiments, R 4 is -OR A . In some embodiments, R 4 is -N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently hydrogen. In some embodiments, one of R 3a and R 3b is independently C 1 -C 6 alkyl. In some embodiments, one of R 3a and R 3b is independently C 2 -C 6 alkenyl. In some embodiments, one of R 3a and R 3b is independently C2-C6 alkynyl. In some embodiments, one of R 3a and R 3b is independently C1-C6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently C 1 -C 6 haloalkyl.
  • one of R 3a and R 3b is independently halo. In some embodiments, one of R 3a and R 3b is independently cyano. In some embodiments, one of R 3a and R 3b is independently -OR A . In some embodiments, one of R 3a and R 3b is independently - N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen. In some embodiments, each of R 3a and R 3b is independently C1-C6 alkyl. In some embodiments, each of R 3a and R 3b is independently C2-C6 alkenyl. In some embodiments, each of R 3a and R 3b is independently C 2 -C 6 alkynyl.
  • each of R 3a and R 3b is independently C 1 -C 6 heteroalkyl. In some embodiments, each of R 3a and R 3b is independently C 1 -C 6 haloalkyl. In some embodiments, each of R 3a and R 3b is independently halo. In some embodiments, each of R 3a and R 3b is independently cyano. In some embodiments, each of R 3a and R 3b is independently -OR A . In some embodiments, each of R 3a and R 3b is independently -N(R B )(R C ).
  • each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R 5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently hydrogen. In some embodiments, one of R 5a and R 5b is independently C1-C6 alkyl. In some embodiments, one of R 5a and R 5b is independently C2-C6 alkenyl. In some embodiments, one of R 5a and R 5b is independently C 2 -C 6 alkynyl.
  • one of R 5a and R 5b is independently C 1 -C 6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently C1-C6 haloalkyl. In some embodiments, one of R 5a and R 5b is independently halo. In some embodiments, one of R5a and R5b is independently cyano. In some embodiments, one of Attorney Docket No.: F2099-7037WO R 5a and R 5b is independently -OR A . In some embodiments, one of R 5a and R 5b is independently - N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen.
  • each of R 5a and R 5b is independently C 1 -C 6 alkyl. In some embodiments, each of R 5a and R 5b is independently C2-C6 alkenyl. In some embodiments, each of R 5a and R 5b is independently C2-C6 alkynyl. In some embodiments, each of R 5a and R 5b is independently C1-C6 heteroalkyl. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 haloalkyl. In some embodiments, each of R 5a and R 5b is independently halo. In some embodiments, each of R 5a and R 5b is independently cyano.
  • each of R 5a and R 5b is independently -OR A . In some embodiments, each of R 5a and R 5b is independently -N(R B )(R C ). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C 1 -C 6 alkyl. In some embodiments, each R A is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each R A is independently hydrogen, C1- C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, each R A is independently hydrogen or C 1 - C6 alkyl.
  • each R A is independently hydrogen.
  • each of R B and R C is independently hydrogen, C1-C6 alkyl or C1- C 6 -heteroalkyl.
  • each of R B and R C is independently hydrogen or C 1 -C 6 alkyl.
  • each of R B and R C is independently hydrogen.
  • R 10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -OR A1 .
  • R 10 is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or C 1 -C 6 haloalkyl.
  • R 10 is C 1 -C 6 alkyl or C 1 -C 6 heteroalkyl. In some embodiments, R 10 is C1-C6 alkyl. In some embodiments, R 10 is C1-C6 heteroalkyl. In some embodiments, R 10 is C1-C6 haloalkyl. In some embodiments, R 10 is halo. In some embodiments, R 10 is cyano. In some embodiments, R 10 is -OR A1 . In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is C 1 -C 6 alkyl. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1.
  • TREM comprises a single 2’,5’-linked nucleotide. In some embodiments, the TREM comprises a plurality of 2’,5’-linked nucleotides. In some Attorney Docket No.: F2099-7037WO embodiments, the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 252’,5’-linked nucleotides. In some embodiments, the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]- [L2].
  • the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]- [L2]. In some embodiments, the 2’,5’-linked nucleotide is present in the [L2]-[DH Domain]- [L3]. In some embodiments, the 2’,5’-linked nucleotide is present in the [L3]-[ACH Domain]. In some embodiments, the 2’,5’-linked nucleotide is present in the [VL Domain]. In some embodiments, the 2’,5’-linked nucleotide is present in the [TH Domain].
  • the 2’,5’-linked nucleotide is present in the [L4]-[ASt Domain2]-[L5].
  • the TREM has a sequence selected from a sequence provided in FIG.1.
  • the TREM comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in Table 1.
  • the TREM comprises a nucleotide sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from the nucleotide sequence of a TREM provided in Table 1, e.g., any one of the TREMs having SEQ ID NO.794-991, 1060, 1062, 1112, 1135, 1153-1158, 1161, 1163, 1166-1170, 1175, 1200, 1207, and 1253-1415 (e.g., any one of the TREMs having SEQ ID NO.794-991, e.g., any one of the TREMs having SEQ ID NO.
  • the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non- naturally occurring modifications compared with a TREM provided in Table 1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications).
  • Table 1 e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications.
  • the TREM comprises the nucleotide sequence of any one of the TREMs having SEQ ID NO.794-991, 1060, 1062, 1112, 1135, 1153- 1158, 1161, 1163, 1166-1170, 1175, 1200, 1207, or 1253-1415 (e.g., any one of the TREMs having SEQ ID NO.794-991.
  • the present disclosure consists of a TREM sequence comprising the 2’,5’- linked nucleotide having a structure of Formula (III-a): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, or N(R B ); each of R 1 , R 2 , and R 4 is independently C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; R 3a is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl,
  • B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). In some embodiments, B is a non-naturally occurring nucleobase. In some embodiments, B comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. In some embodiments, the chemical modification is a nucleotide sugar modification. In some embodiments, the chemical Attorney Docket No.: F2099-7037WO modification is a 2’OMe modification.
  • the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification. In some embodiments, the chemical modification is a 2’MOE modification. In some embodiments, the chemical modification is a 2’-deoxy modification. In some embodiments, the chemical modification is present in the internucleotide region (e.g., a backbone modification). In some embodiments, the chemical modification is a phosphorothioate modification. In some embodiments, B does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(R B ).
  • R 1 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C1-C6 alkyl. In some embodiments, R 1 is C 2 -C 6 alkenyl. In some embodiments, R 1 is C 2 -C 6 alkynyl. In some embodiments, R 1 is C 1 - C6 heteroalkyl. In some embodiments, R 1 is C1-C6 haloalkyl.
  • R 1 is halo. In some embodiments, R 1 is cyano. In some embodiments, R 1 is -OR A . In some embodiments, R 1 is -N(R B )(R C ). In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is C 1 -C 6 alkyl. In some embodiments, R 2 is C 2 -C 6 alkenyl.
  • R 2 is C 2 -C 6 alkynyl. In some embodiments, R 2 is C 1 - C6 heteroalkyl. In some embodiments, R 2 is C1-C6 haloalkyl. In some embodiments, R 2 is halo. In some embodiments, R 2 is cyano. In some embodiments, R 2 is -OR A . In some embodiments, R 2 is -N(R B )(R C ). In some embodiments, R 4 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • R 4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is C 1 -C 6 alkyl. In some embodiments, R 4 is C 2 -C 6 alkenyl. In some embodiments, R 4 is C 2 -C 6 alkynyl. In some embodiments, R 4 is C 1 - C6 heteroalkyl. In some embodiments, R 4 is C1-C6 haloalkyl. In some embodiments, R 4 is halo. In some embodiments, R4 is cyano. In some embodiments, R4 is -ORA.
  • R4 is -N(R B )(R C ). Attorney Docket No.: F2099-7037WO
  • R 3a is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 - C6 haloalkyl, or halo.
  • R 3a is independently hydrogen, C1-C6 alkyl, or C1- C6 heteroalkyl.
  • R 3a is hydrogen.
  • R 3a is C1-C6 alkyl.
  • R 3a is C 2 -C 6 alkenyl.
  • R 3a is C 2 -C 6 alkynyl.
  • R 3a is C1-C6 heteroalkyl. In some embodiments, R 3a is C1-C6 haloalkyl. In some embodiments, R 3a is halo. In some embodiments, R 3a is cyano. In some embodiments, R 3a is - OR A . In some embodiments, R 3a is -N(R B )(R C ).
  • R 3c is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 .
  • R 3c is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl.
  • R 3c is hydrogen.
  • R 3c is C1-C6 alkyl.
  • R 3c is C1-C6 heteroalkyl.
  • each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently hydrogen. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 alkyl. In some embodiments, one of R 5a and R 5b is independently C 2 -C 6 alkenyl. In some embodiments, one of R 5a and R 5b is independently C 2 -C 6 alkynyl.
  • one of R 5a and R 5b is independently C1-C6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 haloalkyl. In some embodiments, one of R 5a and R 5b is independently halo. In some embodiments, one of R 5a and R 5b is independently cyano. In some embodiments, one of R 5a and R 5b is independently -OR A . In some embodiments, one of R 5a and R 5b is independently - N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 alkyl.
  • each of R 5a and R 5b is independently C2-C6 alkenyl. In some embodiments, each of R 5a and R 5b is independently C 2 -C 6 alkynyl. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 heteroalkyl. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 haloalkyl. In some embodiments, each of R 5a and R 5b is independently halo. In some embodiments, each of R5a and R5b is independently cyano. In some embodiments, each of R5a and R5b is independently -OR A .
  • each of R 5a and R 5b is independently -N(R B )(R C ). Attorney Docket No.: F2099-7037WO
  • R’ is hydrogen.
  • R’ is C 1 -C 6 alkyl.
  • each R A is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl.
  • each R A is independently hydrogen, C1- C 6 alkyl, or C 1 -C 6 heteroalkyl.
  • each R A is independently hydrogen or C 1 - C6 alkyl.
  • each R A is independently hydrogen.
  • each of R B and R C is independently hydrogen, C1-C6 alkyl or C1- C 6 -heteroalkyl. In some embodiments, each of R B and R C is independently hydrogen or C 1 -C 6 alkyl. In some embodiments, each of R B and R C is independently hydrogen. In some embodiments, R 10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -OR A1 . In some embodiments, R 10 is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or C 1 -C 6 haloalkyl.
  • R 10 is C 1 -C 6 alkyl or C 1 -C 6 heteroalkyl. In some embodiments, R 10 is C1-C6 alkyl. In some embodiments, R 10 is C1-C6 heteroalkyl. In some embodiments, R 10 is C1-C6 haloalkyl. In some embodiments, R 10 is halo. In some embodiments, R 10 is cyano. In some embodiments, R 10 is -OR A1 . In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is C1-C6 alkyl. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1.
  • the nucleotide within the TREM sequence comprising the 2’,5’- linked nucleotide has a structure of Formula (III-a-i) or Formula (III-a-ii):
  • the present disclosure consists of a TREM sequence comprising the 2’,5’- linked nucleotide having a structure of Formula (III-b): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y and R 4 is independently hydrogen, C1-C6 alkyl, C2-C6 C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and
  • Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(R B ). In some embodiments, R 1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C 1 -C 6 alkyl. In some embodiments, Attorney Docket No.: F2099-7037WO R 1 is C 2 -C 6 alkenyl.
  • R 1 is C 2 -C 6 alkynyl. In some embodiments, R 1 is C 1 - C6 heteroalkyl. In some embodiments, R 1 is C1-C6 haloalkyl. In some embodiments, R 1 is halo. In some embodiments, R 1 is cyano. In some embodiments, R 1 is -OR A . In some embodiments, R 1 is -N(R B )(R C ). In some embodiments, R 2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • R 2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is C 1 -C 6 alkyl. In some embodiments, R 2 is C2-C6 alkenyl. In some embodiments, R 2 is C2-C6 alkynyl. In some embodiments, R 2 is C1- C6 heteroalkyl. In some embodiments, R 2 is C1-C6 haloalkyl. In some embodiments, R 2 is halo. In some embodiments, R 2 is cyano. In some embodiments, R 2 is -OR A . In some embodiments, R 2 is -N(R B )(R C ).
  • R 4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is C 1 -C 6 alkyl. In some embodiments, R 4 is C2-C6 alkenyl. In some embodiments, R 4 is C2-C6 alkynyl. In some embodiments, R 4 is C1- C6 heteroalkyl. In some embodiments, R 4 is C1-C6 haloalkyl. In some embodiments, R 4 is halo.
  • R 4 is cyano. In some embodiments, R 4 is -OR A . In some embodiments, R 4 is -N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently hydrogen. In some embodiments, one of R 3a and R 3b is independently C1-C6 alkyl.
  • one of R 3a and R 3b is independently C2-C6 alkenyl. In some embodiments, one of R 3a and R 3b is independently C 2 -C 6 alkynyl. In some embodiments, one of R 3a and R 3b is independently C 1 -C 6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently C1-C6 haloalkyl. In some embodiments, one of R 3a and R 3b is independently halo. In some embodiments, one of R 3a and R 3b is independently cyano. In some embodiments, one of R 3a and R 3b is independently -OR A .
  • R 3a and R 3b are independently - N(R B )(R C ). I n some embodiments, each of R3a and R3b is independently hydrogen. In some embodiments, each of R 3a and R 3b is independently C 1 -C 6 alkyl. In some embodiments, each of Attorney Docket No.: F2099-7037WO R 3a and R 3b is independently C 2 -C 6 alkenyl. In some embodiments, each of R 3a and R 3b is independently C2-C6 alkynyl. In some embodiments, each of R 3a and R 3b is independently C1-C6 heteroalkyl.
  • each of R 3a and R 3b is independently C1-C6 haloalkyl. In some embodiments, each of R 3a and R 3b is independently halo. In some embodiments, each of R 3a and R 3b is independently cyano. In some embodiments, each of R 3a and R 3b is independently -OR A . In some embodiments, each of R 3a and R 3b is independently -N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently hydrogen. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 alkyl. In some embodiments, one of R 5a and R 5b is independently C 2 -C 6 alkenyl. In some embodiments, one of R 5a and R 5b is independently C2-C6 alkynyl. In some embodiments, one of R 5a and R 5b is independently C1-C6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 haloalkyl.
  • one of R 5a and R 5b is independently halo. In some embodiments, one of R 5a and R 5b is independently cyano. In some embodiments, one of R 5a and R 5b is independently -OR A . In some embodiments, one of R 5a and R 5b is independently - N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen. In some embodiments, each of R 5a and R 5b is independently C1-C6 alkyl. In some embodiments, each of R 5a and R 5b is independently C 2 -C 6 alkenyl. In some embodiments, each of R 5a and R 5b is independently C 2 -C 6 alkynyl.
  • each of R 5a and R 5b is independently C 1 -C 6 heteroalkyl. In some embodiments, each of R 5a and R 5b is independently C1-C6 haloalkyl. In some embodiments, each of R 5a and R 5b is independently halo. In some embodiments, each of R 5a and R 5b is independently cyano. In some embodiments, each of R 5a and R 5b is independently -OR A . In some embodiments, each of R 5a and R 5b is independently -N(R B )(R C ). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C1-C6 alkyl.
  • each R A is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 heteroalkyl. In some embodiments, each R A is independently hydrogen, C 1 - C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each R A is independently hydrogen or C1- C 6 alkyl. In some embodiments, each R A is independently hydrogen. Attorney Docket No.: F2099-7037WO In some embodiments, each of R B and R C is independently hydrogen, C 1 -C 6 alkyl or C 1 - C6-heteroalkyl.
  • each of R B and R C is independently hydrogen or C1-C6 alkyl. In some embodiments, each of R B and R C is independently hydrogen. In some embodiments, R 10 is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, or -OR A1 . In some embodiments, R 10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R 10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R 10 is C 1 -C 6 alkyl.
  • R 10 is C 1 -C 6 heteroalkyl. In some embodiments, R 10 is C1-C6 haloalkyl. In some embodiments, R 10 is halo. In some embodiments, R 10 is cyano. In some embodiments, R 10 is -OR A1 . In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is C 1 -C 6 alkyl. In some embodiments, each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl or C 1 - C6-heteroalkyl. In some embodiments, each of R ia and R ib is independently hydrogen or C1-C6 alkyl.
  • each of R ia and R ib is independently hydrogen. In some embodiments, each of R ii and R iii is independently hydrogen, C 1 -C 6 alkyl, or C 1 - C6 heteroalkyl. In some embodiments, each of R ii and R iii is independently hydrogen or C1-C6 alkyl. In some embodiments, each of R ii and R iii is independently hydrogen. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2.
  • the present disclosure consists of a TREM sequence comprising the 2’,5’-linked nucleotide having a structure of Formula (III-c): a salt, stereoisomer, tautomer, or hydrate R 2 , and R 4 is independently hydrogen, C 1 - C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally Attorney Docket No.: F2099-7037WO substituted with one or more R 10 ; each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, C 2 - C6 alkenyl, C2-C6 alkynyl, C1-C6
  • Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(R B ). In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C1-C6 alkyl. In some embodiments, R 1 is C2-C6 alkenyl. In some embodiments, R 1 is C2-C6 alkynyl.
  • R 1 is C1- C 6 heteroalkyl. In some embodiments, R 1 is C 1 -C 6 haloalkyl. In some embodiments, R 1 is halo. In some embodiments, R 1 is cyano. In some embodiments, R 1 is -OR A . In some embodiments, R 1 is -N(R B )(R C ). In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 2 is hydrogen.
  • R 2 is C1-C6 alkyl. In some embodiments, R2 is C 2 -C 6 alkenyl. In some embodiments, R2 is C 2 -C 6 alkynyl. In some embodiments, R2 is C 1 - C 6 heteroalkyl. In some embodiments, R 2 is C 1 -C 6 haloalkyl. In some embodiments, R 2 is halo. Attorney Docket No.: F2099-7037WO In some embodiments, R 2 is cyano. In some embodiments, R 2 is -OR A . In some embodiments, R 2 is -N(R B )(R C ).
  • R 4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 4 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is C1-C6 alkyl. In some embodiments, R 4 is C2-C6 alkenyl. In some embodiments, R 4 is C2-C6 alkynyl. In some embodiments, R 4 is C1- C 6 heteroalkyl. In some embodiments, R 4 is C 1 -C 6 haloalkyl. In some embodiments, R 4 is halo.
  • R 4 is cyano. In some embodiments, R 4 is -OR A . In some embodiments, R 4 is -N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently hydrogen. In some embodiments, one of R 3a and R 3b is independently C 1 -C 6 alkyl.
  • one of R 3a and R 3b is independently C 2 -C 6 alkenyl. In some embodiments, one of R 3a and R 3b is independently C2-C6 alkynyl. In some embodiments, one of R 3a and R 3b is independently C1-C6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently C 1 -C 6 haloalkyl. In some embodiments, one of R 3a and R 3b is independently halo. In some embodiments, one of R 3a and R 3b is independently cyano. In some embodiments, one of R 3a and R 3b is independently -OR A .
  • one of R 3a and R 3b is independently - N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen. In some embodiments, each of R 3a and R 3b is independently C1-C6 alkyl. In some embodiments, each of R 3a and R 3b is independently C2-C6 alkenyl. In some embodiments, each of R 3a and R 3b is independently C 2 -C 6 alkynyl. In some embodiments, each of R 3a and R 3b is independently C 1 -C 6 heteroalkyl. In some embodiments, each of R 3a and R 3b is independently C 1 -C 6 haloalkyl.
  • each of R 3a and R 3b is independently halo. In some embodiments, each of R 3a and R 3b is independently cyano. In some embodiments, each of R 3a and R 3b is independently -OR A . In some embodiments, each of R 3a and R 3b is independently -N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl.
  • one of R 5a Attorney Docket No.: F2099-7037WO and R 5b is independently hydrogen. In some embodiments, one of R 5a and R 5b is independently C1-C6 alkyl. In some embodiments, one of R 5a and R 5b is independently C2-C6 alkenyl. In some embodiments, one of R 5a and R 5b is independently C2-C6 alkynyl. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently C1-C6 haloalkyl. In some embodiments, one of R 5a and R 5b is independently halo.
  • one of R 5a and R 5b is independently cyano. In some embodiments, one of R 5a and R 5b is independently -OR A . In some embodiments, one of R 5a and R 5b is independently - N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 alkyl. In some embodiments, each of R 5a and R 5b is independently C 2 -C 6 alkenyl. In some embodiments, each of R 5a and R 5b is independently C2-C6 alkynyl.
  • each of R 5a and R 5b is independently C1-C6 heteroalkyl. In some embodiments, each of R 5a and R 5b is independently C1-C6 haloalkyl. In some embodiments, each of R 5a and R 5b is independently halo. In some embodiments, each of R 5a and R 5b is independently cyano. In some embodiments, each of R 5a and R 5b is independently -OR A . In some embodiments, each of R 5a and R 5b is independently -N(R B )(R C ). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C 1 -C 6 alkyl.
  • each R A is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each R A is independently hydrogen, C1- C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, each R A is independently hydrogen or C 1 - C 6 alkyl. In some embodiments, each R A is independently hydrogen. In some embodiments, each of R B and R C is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of R B and R C is independently hydrogen or C1-C6 alkyl.
  • each of R B and R C is independently hydrogen.
  • R 10 is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, or -OR A1 .
  • R 10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl.
  • R 10 is C 1 -C 6 alkyl or C 1 -C 6 heteroalkyl.
  • R 10 is C 1 -C 6 alkyl.
  • R 10 is C 1 -C 6 heteroalkyl.
  • R 10 is C1-C6 haloalkyl. In some embodiments, R 10 is halo. In some embodiments, R 10 is cyano. In some embodiments, R 10 is -OR A1 . In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is C 1 -C 6 alkyl. Attorney Docket No.: F2099-7037WO In some embodiments, each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl or C 1 - C6-heteroalkyl. In some embodiments, each of R ia and R ib is independently hydrogen or C1-C6 alkyl.
  • each of R ia and R ib is independently hydrogen.
  • R ii is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl.
  • R ii is hydrogen or C1-C6 alkyl.
  • R ii is hydrogen.
  • R iii is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl.
  • R iii is hydrogen or C 1 -C 6 alkyl.
  • R iii is hydrogen.
  • each of x and y is independently 0, 1, or 2; In some embodiments, x is 0.
  • the present disclosure consists of a TREM sequence comprising the 2’,5’-linked nucleotide having a structure of Formula (III-d): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y and R 4 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 3a and R 3b is independently hydrogen,
  • Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(R B ). In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C1-C6 alkyl. In some embodiments, R 1 is C2-C6 alkenyl. In some embodiments, R 1 is C2-C6 alkynyl.
  • R 1 is C1- C 6 heteroalkyl. In some embodiments, R 1 is C 1 -C 6 haloalkyl. In some embodiments, R 1 is halo. In some embodiments, R 1 is cyano. In some embodiments, R 1 is -OR A . In some embodiments, R 1 is -N(R B )(R C ). In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 2 is hydrogen.
  • R 2 is C1-C6 alkyl. In some embodiments, R 2 is C 2 -C 6 alkenyl. In some embodiments, R 2 is C 2 -C 6 alkynyl. In some embodiments, R 2 is C 1 - C 6 heteroalkyl. In some embodiments, R 2 is C 1 -C 6 haloalkyl. In some embodiments, R 2 is halo. In some embodiments, R 2 is cyano. In some embodiments, R 2 is -OR A . In some embodiments, R 2 is -N(R B )(R C ).
  • R 4 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 4 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is C1-C6 alkyl. In some embodiments, R 4 is C 2 -C 6 alkenyl. In some embodiments, R 4 is C 2 -C 6 alkynyl. In some embodiments, R 4 is C 1 - C 6 heteroalkyl. In some embodiments, R 4 is C 1 -C 6 haloalkyl.
  • R 4 is halo. In some embodiments, R 4 is cyano. In some embodiments, R 4 is -OR A . In some embodiments, R 4 is -N(RB)(RC). Attorney Docket No.: F2099-7037WO
  • each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently hydrogen.
  • one of R 3a and R 3b is independently C1-C6 alkyl. In some embodiments, one of R 3a and R 3b is independently C2-C6 alkenyl. In some embodiments, one of R 3a and R 3b is independently C2-C6 alkynyl. In some embodiments, one of R 3a and R 3b is independently C 1 -C 6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently C1-C6 haloalkyl. In some embodiments, one of R 3a and R 3b is independently halo. In some embodiments, one of R 3a and R 3b is independently cyano.
  • one of R 3a and R 3b is independently -OR A . In some embodiments, one of R 3a and R 3b is independently - N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen. In some embodiments, each of R 3a and R 3b is independently C1-C6 alkyl. In some embodiments, each of R 3a and R 3b is independently C 2 -C 6 alkenyl. In some embodiments, each of R 3a and R 3b is independently C2-C6 alkynyl. In some embodiments, each of R 3a and R 3b is independently C1-C6 heteroalkyl.
  • each of R 3a and R 3b is independently C1-C6 haloalkyl. In some embodiments, each of R 3a and R 3b is independently halo. In some embodiments, each of R 3a and R 3b is independently cyano. In some embodiments, each of R 3a and R 3b is independently -OR A . In some embodiments, each of R 3a and R 3b is independently -N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently hydrogen. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 alkyl. In some embodiments, one of R 5a and R 5b is independently C 2 -C 6 alkenyl. In some embodiments, one of R 5a and R 5b is independently C 2 -C 6 alkynyl. In some embodiments, one of R 5a and R 5b is independently C1-C6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 haloalkyl.
  • one of R 5a and R 5b is independently halo. In some embodiments, one of R 5a and R 5b is independently cyano. In some embodiments, one of R 5a and R 5b is independently -OR A . In some embodiments, one of R 5a and R 5b is independently - N(R B )(R C ). Attorney Docket No.: F2099-7037WO In some embodiments, each of R 5a and R 5b is independently hydrogen. In some embodiments, each of R 5a and R 5b is independently C1-C6 alkyl. In some embodiments, each of R 5a and R 5b is independently C2-C6 alkenyl.
  • each of R 5a and R 5b is independently C 2 -C 6 alkynyl. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 heteroalkyl. In some embodiments, each of R 5a and R 5b is independently C1-C6 haloalkyl. In some embodiments, each of R 5a and R 5b is independently halo. In some embodiments, each of R 5a and R 5b is independently cyano. In some embodiments, each of R 5a and R 5b is independently -OR A . In some embodiments, each of R 5a and R 5b is independently -N(R B )(R C ). In some embodiments, R’ is hydrogen.
  • R’ is C1-C6 alkyl.
  • each R A is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 heteroalkyl.
  • each R A is independently hydrogen, C 1 - C6 alkyl, or C1-C6 heteroalkyl.
  • each R A is independently hydrogen or C1- C6 alkyl.
  • each R A is independently hydrogen.
  • each of R B and R C is independently hydrogen, C 1 -C 6 alkyl or C 1 - C6-heteroalkyl.
  • each of R B and R C is independently hydrogen or C1-C6 alkyl. In some embodiments, each of R B and R C is independently hydrogen. In some embodiments, R 10 is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, or -OR A1 . In some embodiments, R 10 is C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or C 1 -C 6 haloalkyl. In some embodiments, R 10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R 10 is C 1 -C 6 alkyl.
  • R 10 is C 1 -C 6 heteroalkyl. In some embodiments, R 10 is C 1 -C 6 haloalkyl. In some embodiments, R 10 is halo. In some embodiments, R 10 is cyano. In some embodiments, R 10 is -OR A1 . In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is C1-C6 alkyl. In some embodiments, each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl or C 1 - C 6 -heteroalkyl. In some embodiments, each of R ia and R ib is independently hydrogen or C 1 -C 6 alkyl.
  • each of R ia and R ib is independently hydrogen. In some embodiments, each of R ii and R iii is independently hydrogen, C 1 -C 6 alkyl, or C 1 - C 6 heteroalkyl. In some embodiments, each of R ii and R iii is independently hydrogen or C 1 -C 6 alkyl. In some embodiments, each of R ii and R iii is independently hydrogen.
  • each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0.
  • y is 1. In some embodiments, y is 2.
  • the present disclosure consists of a TREM sequence comprising the 2’,5’-linked nucleotide having a structure of Formula (III-e): salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y is and R 4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-
  • Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(R B ). In some embodiments, R 1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R 1 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C1-C6 alkyl. In some embodiments, R 1 is C2-C6 alkenyl. In some embodiments, R 1 is C2-C6 alkynyl.
  • R 1 is C1- C 6 heteroalkyl. In some embodiments, R 1 is C 1 -C 6 haloalkyl. In some embodiments, R 1 is halo. In some embodiments, R 1 is cyano. In some embodiments, R 1 is -OR A . In some embodiments, R 1 is -N(R B )(R C ). In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 2 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 2 is hydrogen.
  • R 2 is C1-C6 alkyl. In some embodiments, R 2 is C2-C6 alkenyl. In some embodiments, R 2 is C2-C6 alkynyl. In some embodiments, R 2 is C1- C 6 heteroalkyl. In some embodiments, R 2 is C 1 -C 6 haloalkyl. In some embodiments, R 2 is halo. In some embodiments, R 2 is cyano. In some embodiments, R 2 is -OR A . In some embodiments, R 2 is -N(R B )(R C ).
  • R 4 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, R 4 is hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, R 4 is hydrogen. In some embodiments, R 4 is C1-C6 alkyl. In some embodiments, R 4 is C 2 -C 6 alkenyl. In some embodiments, R 4 is C 2 -C 6 alkynyl. In some embodiments, R 4 is C 1 - C 6 heteroalkyl. In some embodiments, R 4 is C 1 -C 6 haloalkyl.
  • R 4 is halo. In some embodiments, R 4 is cyano. In some embodiments, R 4 is -OR A . In some embodiments, R 4 is -N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R 3a and R 3b is independently hydrogen.
  • one of R 3a and R 3b is independently C 1 -C 6 alkyl. In some embodiments, one of R 3a and R 3b is independently C 2 -C 6 alkenyl. In some embodiments, one of R 3a and R 3b is independently C2-C6 alkynyl. In some embodiments, one of R3a and R3b is independently C 1 -C 6 heteroalkyl. In some embodiments, one of R3a and R 3b is independently C 1 -C 6 haloalkyl. In some embodiments, one of R 3a and R 3b is independently halo. Attorney Docket No.: F2099-7037WO In some embodiments, one of R 3a and R 3b is independently cyano.
  • one of R 3a and R 3b is independently -OR A . In some embodiments, one of R 3a and R 3b is independently - N(R B )(R C ). In some embodiments, each of R 3a and R 3b is independently hydrogen. In some embodiments, each of R 3a and R 3b is independently C1-C6 alkyl. In some embodiments, each of R 3a and R 3b is independently C2-C6 alkenyl. In some embodiments, each of R 3a and R 3b is independently C 2 -C 6 alkynyl. In some embodiments, each of R 3a and R 3b is independently C 1 -C 6 heteroalkyl.
  • each of R 3a and R 3b is independently C1-C6 haloalkyl. In some embodiments, each of R 3a and R 3b is independently halo. In some embodiments, each of R 3a and R 3b is independently cyano. In some embodiments, each of R 3a and R 3b is independently -OR A . In some embodiments, each of R 3a and R 3b is independently -N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • each of R 5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, or C 1 -C 6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently hydrogen. In some embodiments, one of R 5a and R 5b is independently C1-C6 alkyl. In some embodiments, one of R 5a and R 5b is independently C2-C6 alkenyl. In some embodiments, one of R 5a and R 5b is independently C 2 -C 6 alkynyl. In some embodiments, one of R 5a and R 5b is independently C 1 -C 6 heteroalkyl. In some embodiments, one of R 5a and R 5b is independently C1-C6 haloalkyl.
  • one of R 5a and R 5b is independently halo. In some embodiments, one of R 5a and R 5b is independently cyano. In some embodiments, one of R 5a and R 5b is independently -OR A . In some embodiments, one of R 5a and R 5b is independently - N(R B )(R C ). In some embodiments, each of R 5a and R 5b is independently hydrogen. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 alkyl. In some embodiments, each of R 5a and R 5b is independently C 2 -C 6 alkenyl. In some embodiments, each of R 5a and R 5b is independently C2-C6 alkynyl.
  • each of R 5a and R 5b is independently C1-C6 heteroalkyl. In some embodiments, each of R 5a and R 5b is independently C 1 -C 6 haloalkyl. In some embodiments, each of R 5a and R 5b is independently halo. In some embodiments, each of R 5a and R 5b is independently cyano. In some embodiments, each of R 5a and R 5b is independently -ORA. In some embodiments, each of R5a and R5b is independently -N(RB)(RC). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C 1 -C 6 alkyl.
  • each R A is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each R A is independently hydrogen, C1- C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each R A is independently hydrogen or C1- C 6 alkyl. In some embodiments, each R A is independently hydrogen. In some embodiments, each of R B and R C is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl.
  • each of R B and R C is independently hydrogen or C1-C6 alkyl. In some embodiments, each of R B and R C is independently hydrogen. In some embodiments, R 10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -OR A1 . In some embodiments, R 10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R 10 is C 1 -C 6 alkyl or C 1 -C 6 heteroalkyl. In some embodiments, R 10 is C 1 -C 6 alkyl.
  • R 10 is C 1 -C 6 heteroalkyl. In some embodiments, R 10 is C1-C6 haloalkyl. In some embodiments, R 10 is halo. In some embodiments, R 10 is cyano. In some embodiments, R 10 is -OR A1 . In some embodiments, R A1 is hydrogen. In some embodiments, R A1 is C 1 -C 6 alkyl. In some embodiments, R i is hydrogen, C1-C6 alkyl or C1-C6-heteroalkyl. In some embodiments, R i is hydrogen or C1-C6 alkyl. In some embodiments, R i is hydrogen.
  • each of R ii and R iii is independently hydrogen, C 1 -C 6 alkyl, or C 1 - C 6 heteroalkyl. In some embodiments, each of R ii and R iii is independently hydrogen or C 1 -C 6 alkyl. In some embodiments, each of R ii and R iii is independently hydrogen. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2.
  • the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease.
  • a TREM may comprise multiple 2’,5’-linked nucleotides independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].
  • the TREM comprises one 2’,5’-linked nucleotide.
  • the TREM contains a 2’,5’-linked nucleotide located in the [ASt Domain1].
  • the TREM contains a 2’,5’-linked nucleotide located in the [DH Domain].
  • the TREM contains a 2’,5’-linked nucleotide located in the [ACH Domain]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [VL Attorney Docket No.: F2099-7037WO Domain]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [TH Domain]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [Ast2 Domain]. In an embodiment, the 2’,5’-linked nucleotide is located at the nucleotide position 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of the TREM.
  • the 2’,5’-linked nucleotide is located at the nucleotide position 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or 75 of the TREM.
  • the TREM contains two or more 2’,5’-linked nucleotides. In an embodiment, the TREM contains two 2’,5’-linked nucleotides.
  • the TREM contains two 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [ACH Domain]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [DH Domain]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [DH Domain] and the [TH Domain]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [TH Domain].
  • the TREM contains two 2’,5’-linked nucleotides each independently located in the [ASt Domain 1]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located between nucleotide positions 30 and 40. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located between nucleotide positions 12 and 25. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located between nucleotide positions 30 and 40. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at nucleotide positions 31 and 32.
  • the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 31 and 34. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 31 and 35. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 31 and 37. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 32 and 34.
  • Attorney Docket No.: F2099-7037WO A TREM may comprise three 2’,5’-linked nucleotides.
  • the TREM comprises three 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]..
  • the TREM contains three 2’,5’-linked nucleotides each independently located between nucleotide positions 9 and 60.
  • a TREM may comprise four 2’,5’-linked nucleotides.
  • the TREM comprises four 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].
  • a TREM may comprise five 2’,5’-linked nucleotides.
  • the TREM comprises five 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].
  • a TREM may comprise six 2’,5’-linked nucleotides.
  • the TREM comprises six 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].
  • the TREM contains a 2’,5’-linked nucleotide at nucleotide position 49.
  • a TREM may contain several non-naturally occurring modifications in addition to 2’,5’- linked nucleotides.
  • a TREM may comprise a non-naturally occurring modification in each of the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].
  • the TREM comprises four 2’,5’-linked nucleotides independently located in the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].
  • the TREM comprises four 2’,5’-linked nucleotides independently located in at nucleotide positions 43, 45, 54, and 65.
  • the TREM comprises five 2’,5’-linked nucleotides, in addition to 24 2’-O-methyl modifications. In an embodiment, the TREM comprises five 2’,5’-linked nucleotides, in addition to 142’-fluoro modifications. In an embodiment, the TREM comprises five locked nucleic acids, in addition to one 2’-O-methoxyethyl modification. In an embodiment, the TREM comprises five locked nucleic acids, in addition to nine phosphorothioate modifications.
  • the TREM comprises five locked nucleic acids independently located in the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2], 242’-O-methyl modifications independently located in the Attorney Docket No.: F2099-7037WO [ASt Domain 1], [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2], 142’-fluoro modifications independently located in the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2], one 2’-O- methoxyethyl modification located in the [ACH Domain], and nine phosphorothioate modifications independently located in the [ASt Domain1], [DH Domain], the [ACH Domain], the [TH Domain], and the [ASt Domain2].
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.176. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.193. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.202. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.204. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.207. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.216.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.217. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.218. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.219. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.220. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.222. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.224.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.226. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.227. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.228. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.229. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.232. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.233.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.238. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.239. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.240. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.241. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.242. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.243.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.244. In some Attorney Docket No.: F2099-7037WO embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.245. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.246. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.247. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.248.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.249. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.250. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.251. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.252. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.253. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.254.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.255. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.256. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.257. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.258. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.259. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.260.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.262. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.263. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.264. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.265. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.266. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.267.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.269. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.270. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.272. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.273. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.274. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.275.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.276. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.277. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.278. In some Attorney Docket No.: F2099-7037WO embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.279. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.281.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.282. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.283. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.284. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.285. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.286. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.287.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.288. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.289. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.290. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.291. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.292. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.293.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.294. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.295. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.296. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.297. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.298. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.299.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.301. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.302. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.304. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.305. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.307. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.309.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.310. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.311. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.312. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.313. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.315.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.316. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.317. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.318. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.320. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.321.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.322. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.323. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.324. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.325. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.326. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.327.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.328. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.329. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.330. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.331. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.332. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.333.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.334. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.335. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.336. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.337. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.338. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.339.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.340. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.341. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.342. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.343. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.346. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.357.
  • the TREM containing a 2’,5’-linked nucleotide is TREM No.361.
  • TREMS CONTAINING ADDITIONAL MODIFICATIONS Attorney Docket No.: F2099-7037WO Described herein are TREMs comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, and at least one additional non-naturally occurring modifications.
  • the TREM comprises a locked nucleic acid moiety and a 2’,5’-linked nucleotide.
  • the TREM comprises a locked nucleic acid moiety and a 2’,5’-linked nucleotide, as well as an additional non-naturally occurring modification.
  • the TREM comprises a single LNA moiety or 2’,5’-linked nucleotide. In some embodiments, the TREM comprises a plurality of LNA moieties and 2’,5’- linked nucleotides. In some embodiments, the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 LNA moieties or 2’,5’-linked nucleotides. In some embodiments, the TREM comprises at least one LNA moiety and at least one 2’,5’-linked nucleotide. In some embodiments, the TREM comprises one LNA moiety and one 2’,5’-linked nucleotide.
  • the TREM comprising an LNA moiety and a 2’,5’-linked nucleotide is a TREM having any one of the SEQ ID Nos: 1060, 1062, 1112, 1135, 1153-1158, 1161, 1163, 1166-1170, 1175, 1200, 1207, 1357, 1364, 1378-1393, and 1397-1398.
  • the TREM comprises two LNA moieties and at least one 2’,5’- linked nucleotide.
  • the TREM comprises two LNA moieties and one 2’,5’- linked nucleotide.
  • the TREM comprises two LNA moieties and two 2’,5’- linked nucleotides.
  • the TREM comprises two LNA moieties and three 2’,5’-linked nucleotides. In some embodiments, the TREM comprises two LNA moieties and four 2’,5’-linked nucleotides. In some embodiments, the TREM comprises three LNA moieties and at least one 2’,5’- linked nucleotide. In some embodiments, the TREM comprises three LNA moieties and one 2’,5’-linked nucleotide. In some embodiments, the TREM comprises three LNA moieties and two 2’,5’-linked nucleotides. In some embodiments, the TREM comprises three LNA moieties and three 2’,5’-linked nucleotides.
  • the TREM comprises four LNA moieties and at least one 2’,5’- linked nucleotide. In some embodiments, the TREM comprises four LNA moieties and one 2’,5’-linked nucleotide. In some embodiments, the TREM comprises four LNA moieties and two 2’,5’-linked nucleotides.
  • the TREM comprising both LNA moieties and 2’,5’-linked nucleotides also comprises at least one additional non-naturally occurring modification (e.g., 2’- Attorney Docket No.: F2099-7037WO ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’- deoxy, or phosphorothiorate modifications).
  • additional non-naturally occurring modification e.g., 2’- Attorney Docket No.: F2099-7037WO ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’- deoxy, or phosphorothiorate modifications.
  • the TREM comprising an LNA moiety and a 2’,5’-linked nucleotide comprises at least 2, 3, 4, 5, 6, 7, 8.9, 10, or more additional non-naturally occurring modifications (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’- halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications).
  • the TREM comprises one LNA moiety, one 2’,5’-linked nucleotide, and at least one 5-methylcytosine.
  • the TREM comprises one LNA moiety, one 2’,5’-linked nucleotide, and one 5-methylcytosine.
  • the TREM comprises one LNA moiety, one 2’,5’-linked nucleotide, and at least one 2’-moe.
  • the present disclosure is a method of treating a subject having a disease or disorder associated with a PTC comprising administering to the subject a TREM, TREM core fragment, or TREM fragment described herein (e.g., a TREM described herein), thereby treating the subject having the disease or disorder.
  • the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease.
  • the TREM comprising an LNA moiety or a 2’,5’-linked nucleotide comprises a sequence corresponding to the nucleotide sequence of SEQ ID NO.625.
  • the TREM comprising an LNA moiety or a 2’,5’-linked nucleotide comprises a sequence corresponding to the nucleotide sequence of SEQ ID NO.1416.
  • TREM, TREM core fragment and TREM fragment fusions In an embodiment, a TREM, a TREM core fragment or a TREM fragment disclosed herein comprises an additional moiety, e.g., a fusion moiety.
  • the fusion moiety can be used for purification, to alter folding of the TREM, TREM core fragment or TREM fragment, or as a targeting moiety.
  • the fusion moiety can comprise a tag, a linker, can be cleavable or can include a binding site for an enzyme.
  • the fusion moiety can be disposed at the N terminal of the TREM or at the C terminal of the TREM, TREM core fragment or TREM fragment.
  • the fusion moiety can be encoded by the same or different nucleic acid molecule that encodes the TREM, TREM core fragment or TREM fragment.
  • a TREM disclosed herein comprises a consensus sequence provided herein.
  • a TREM disclosed herein comprises a consensus sequence of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids and Formula I corresponds to all species.
  • a TREM disclosed herein comprises a consensus sequence of Formula II ZZZ , wherein ZZZ indicates any of the twenty amino acids and Formula II corresponds to mammals.
  • a TREM disclosed herein comprises a consensus sequence of Formula III ZZZ , wherein ZZZ indicates any of the twenty amino acids and Formula III corresponds to humans.
  • ZZZ indicates any of the twenty amino acids: alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine.
  • a TREM disclosed herein comprises a property selected from the following: a) under physiological conditions residue R 0 forms a linker region, e.g., a Linker 1 region; b) under physiological conditions residues R1-R2-R3-R4 -R5-R6-R7 and residues R65-R66- R67-R68-R69-R70-R71 form a stem region, e.g., an AStD stem region; c) under physiological conditions residues R 8 -R 9 forms a linker region, e.g., a Linker 2 region; d) under physiological conditions residues -R10-R11-R12-R13-R14 R15-R16-R17-R18-R19-R20- R 21 -R 22 -R 23 -R 24 -R 25 -R 26 -R 27 -R 28 form a stem-loop region, e.g., a D arm Region; e) under physiological conditions residue residues residue R
  • a TREM disclosed herein comprises the sequence of Formula IALA (SEQ ID NO: 562), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-
  • a TREM disclosed herein comprises the sequence of Formula II ALA (SEQ ID NO: 563), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62
  • a TREM disclosed herein comprises the sequence of Formula IIIALA (SEQ ID NO: 564), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-
  • a TREM disclosed herein comprises the sequence of Formula I ARG (SEQ ID NO: 565), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-
  • a TREM disclosed herein comprises the sequence of Formula II ARG (SEQ ID NO: 566), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R
  • a TREM disclosed herein comprises the sequence of Formula III ARG (SEQ ID NO: 567), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises the sequence of Formula I ASN (SEQ ID NO: 568), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R
  • a TREM disclosed herein comprises the sequence of Formula II ASN (SEQ ID NO: 569), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R
  • a TREM disclosed herein comprises the sequence of Formula III ASN (SEQ ID NO: 570), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R
  • a TREM disclosed herein comprises the sequence of Formula I ASP (SEQ ID NO: 571), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55
  • a TREM disclosed herein comprises the sequence of Formula II ASP (SEQ ID NO: 572), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60
  • a TREM disclosed herein comprises the sequence of Formula III ASP (SEQ ID NO: 573), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises the sequence of Formula I CYS (SEQ ID NO: 574), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-
  • a TREM disclosed herein comprises the sequence of Formula II CYS (SEQ ID NO: 575), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60
  • a TREM disclosed herein comprises the sequence of Formula III CYS (SEQ ID NO: 576), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R
  • a TREM disclosed herein comprises the sequence of Formula I GLN (SEQ ID NO: 577), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-
  • a TREM disclosed herein comprises the sequence of Formula II GLN (SEQ ID NO: 578), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R
  • a TREM disclosed herein comprises the sequence of Formula III GLN (SEQ ID NO: 579), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -
  • a TREM disclosed herein comprises the sequence of Formula I GLU (SEQ ID NO: 580), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-
  • a TREM disclosed herein comprises the sequence of Formula II GLU (SEQ ID NO: 581), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-
  • a TREM disclosed herein comprises the sequence of Formula III GLU (SEQ ID NO: 582), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R
  • a TREM disclosed herein comprises the sequence of Formula I GLY (SEQ ID NO: 583), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R
  • a TREM disclosed herein comprises the sequence of Formula II GLY (SEQ ID NO: 584), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-
  • a TREM disclosed herein comprises the sequence of Formula III GLY (SEQ ID NO: 585), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises the sequence of Formula I HIS (SEQ ID NO: 586), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R
  • a TREM disclosed herein comprises the sequence of Formula II HIS (SEQ ID NO: 587), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R
  • a TREM disclosed herein comprises the sequence of Formula III HIS (SEQ ID NO: 588), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises the sequence of Formula I ILE (SEQ ID NO: 589), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R
  • a TREM disclosed herein comprises the sequence of Formula II ILE (SEQ ID NO: 590), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises the sequence of Formula III ILE (SEQ ID NO: 591), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R
  • a TREM disclosed herein comprises the sequence of Formula I MET (SEQ ID NO: 592), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54
  • a TREM disclosed herein comprises the sequence of Formula II MET (SEQ ID NO: 593), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62
  • a TREM disclosed herein comprises the sequence of Formula III MET (SEQ ID NO: 594), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66
  • a TREM disclosed herein comprises the sequence of Formula I LEU (SEQ ID NO: 595), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R
  • a TREM disclosed herein comprises the sequence of Formula II LEU (SEQ ID NO: 596), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R
  • a TREM disclosed herein comprises the sequence of Formula III LEU (SEQ ID NO: 597), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66
  • a TREM disclosed herein comprises the sequence of Formula I LYS (SEQ ID NO: 598), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R
  • a TREM disclosed herein comprises the sequence of Formula II LYS (SEQ ID NO: 599), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-
  • a TREM disclosed herein comprises the sequence of Formula III LYS (SEQ ID NO: 600), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R
  • a TREM disclosed herein comprises the sequence of Formula I PHE (SEQ ID NO: 601), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54
  • a TREM disclosed herein comprises the sequence of Formula II PHE (SEQ ID NO: 602), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66
  • a TREM disclosed herein comprises the sequence of Formula III PHE (SEQ ID NO: 603), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises the sequence of Formula I PRO (SEQ ID NO: 604), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63
  • a TREM disclosed herein comprises the sequence of Formula II PRO (SEQ ID NO: 605), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R
  • a TREM disclosed herein comprises the sequence of Formula III PRO (SEQ ID NO: 606), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-
  • a TREM disclosed herein comprises the sequence of Formula I SER (SEQ ID NO: 607), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63
  • a TREM disclosed herein comprises the sequence of Formula III SER (SEQ ID NO: 609), R 0 - R 1 - R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R
  • a TREM disclosed herein comprises the sequence of Formula I THR (SEQ ID NO: 610), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R
  • a TREM disclosed herein comprises the sequence of Formula II THR (SEQ ID NO: 611), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60
  • a TREM disclosed herein comprises the sequence of Formula III THR (SEQ ID NO: 612), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66
  • a TREM disclosed herein comprises the sequence of Formula I TRP (SEQ ID NO: 613), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R
  • a TREM disclosed herein comprises the sequence of Formula II TRP (SEQ ID NO: 614), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises the sequence of Formula III TRP (SEQ ID NO: 615), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66
  • a TREM disclosed herein comprises the sequence of Formula I TYR (SEQ ID NO: 616), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R
  • a TREM disclosed herein comprises the sequence of Formula II TYR (SEQ ID NO: 617), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60
  • a TREM disclosed herein comprises the sequence of Formula III TYR (SEQ ID NO: 618), R 0 - R 1 -R 2 - R 3 -R 4 -R 5 -R 6 -R 7 -R 8 -R 9 -R 10 -R 11 -R 12 -R 13 -R 14 -R 15 -R 16 -R 17 -R 18 -R 19 -R 20 -R 21 -R 22 - R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R
  • a TREM disclosed herein comprises the sequence of Formula I VAL (SEQ ID NO: 619), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55
  • a TREM disclosed herein comprises the sequence of Formula II VAL (SEQ ID NO: 620), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45 - R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60
  • a TREM disclosed herein comprises the sequence of Formula III VAL (SEQ ID NO: 621), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R 46 - [R 47 ] x -R 48 -R 49 -R 50 -R 51 -R 52 -R 53 -R 54 -R 55 -R 56 -R 57 -R 58 -R 59 -R 60 -R 61 -R 62 -R 63 -R 64 -R 65
  • a TREM disclosed herein comprises a variable region at position R 47 .
  • the variable region is 1-271 ribonucleotides in length (e.g.1-250, 1-225, 1- 200, 1-175, 1-150, 1-125, 1-100, 1-75, 1-50, 1-40, 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 10-271, 20-271, 30- 271, 40-271, 50-271, 60-271, 70-271, 80-271, 100-271, 125-271, 150-271, 175-271, 200-271, 225-271, 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, 30, 40, 50, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200
  • variable region comprises any one, all or a combination of Adenine, Cytosine, Guanine or Uracil.
  • the variable region comprises a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 4, e.g., any one of SEQ ID NOs: 452-561 disclosed in Table 4.
  • Table 4 Exemplary variable region sequences.
  • Evaluation A 1. The candidate sequence is aligned with each of the consensus sequences in Tables 9 and 10.
  • the consensus sequence(s) having the most positions aligned (and which has at least 60% of the positions of the candidate sequence aligned) is selected.
  • the alignment is performed as is follows.
  • the candidate sequence and an isodecoder consensus sequence from Tables 10A-10B are aligned based on a global pairwise alignment calculated with the Needleman–Wunsch algorithm when run with match scores from Table 11, a mismatch penalty of -1, a gap opening penalty of -1, and a gap extension penalty of -0.5, and no penalty for end gaps.
  • the alignment with the highest overall alignment score is then used to determine the percent similarity between the candidate and the consensus sequence by counting the number of matched positions in the alignment, dividing it by the larger of the number of non- N bases in the candidate sequence or the consensus sequence, and multiplying the result by 100.
  • the percent similarity is the largest percent similarity calculated from the tied alignments. This process is repeated for the candidate sequence with each of the remaining Attorney Docket No.: F2099-7037WO isodecoder consensus sequences in Tables 10A-10B, and the alignment resulting in the greatest percent similarity is selected.
  • this alignment has a percent similarity equal to or greater than 60%, it is considered a valid alignment and used to relate positions in the candidate sequence to those in the consensus sequence, otherwise the candidate sequence is considered to have not aligned to any of the isodecoder consensus sequences. If there is a tie at this point, all tied consensus sequences are taken forward to step 2 in the analysis. 2.
  • the selected consensus sequence(s) from step 1 one determines the consensus sequence position number that aligns with the selected position (e.g., a modified position) in the candidate sequence.
  • step 5 If there were tied consensus sequences from step one, and they give different position numbers in this step 2, then all such position numbers are taken forward to step 5. 3.
  • the reference sequence is aligned with the consensus sequence chosen in step 1. The alignment is performed as described in step 1. 4. From the alignment in step 3, one determines the consensus sequence position number that aligns with the selected position (e.g., a modified position) in the reference sequence. One then assigns the position number of the aligned position in the consensus sequence to the selected position in the reference sequence, in other words, the selected position in the reference sequence is numbered according to the numbering of the consensus sequence. If there is a tie at this point, all tied consensus sequences are taken forward to step 5 in the analysis. 5.
  • Evaluation B The reference sequence (e.g., a TREM sequence described herein) and the candidate sequence are aligned with one another.
  • the alignment is performed as follows.
  • the reference sequence and the candidate sequence are aligned based on a global pairwise alignment calculated with the Needleman–Wunsch algorithm when run with match scores from Table 11, a mismatch penalty of -1, a gap opening penalty of -1, and a gap extension Attorney Docket No.: F2099-7037WO penalty of -0.5, and no penalty for end gaps.
  • the alignment with the highest overall alignment score is then used to determine the percent similarity between the candidate and reference sequence by counting the number of matched based in the alignment, dividing it by the larger of the number of non-N bases in the candidate or reference sequence, and multiplying the result by 100.
  • the percent similarity is the largest percent similarity calculated from the tied alignments. If this alignment has a percent similarity equal to or greater than 60%, it is considered a valid alignment and used to relate positions in the candidate sequence to those in the reference sequence, otherwise the candidate sequence is considered to have not aligned to the reference sequence.
  • the candidate sequence is assigned a nucleotide position number according to the comprehensive tRNA numbering system (CtNS), also referred to as the tRNAviz method (e.g., as described in Lin et al., Nucleic Acids Research, 47:W1, pages W542-W547, 2 July 2019), which serves as a global numbering system for tRNA molecules.
  • CtNS comprehensive tRNA numbering system
  • the alignment is performed as follows. 1.
  • the candidate sequence is assigned a nucleotide position according to the tRNAviz method.
  • the numbering for the closest sequence in the database is obtained. For example, if a TREM differs at any given nucleotide position from a sequence in the database, the numbering for the tRNA having the wildtype sequence at said given nucleotide position is used. 2.
  • the reference sequence is assigned a nucleotide position according to the method described in 1. 3. If a value for a position number determined for the reference sequence in step 1 is the same as the value for the position number determined for the candidate sequence in step 2, the positions are defined as corresponding. If the selected position in the reference sequence and the candidate sequence are found to be corresponding in at least one of Evaluations A, B, and C, the positions correspond.
  • Acid Anticodon Consensus sequence C C C T G C
  • a point mutation in the open reading frame (ORF) of a gene which creates a premature stop codon (PTC) can result in altered expression and/or activity of a polypeptide encoded by the gene.
  • Table 15 provides single mutations in codons encoding amino acids which can result in a stop codon.
  • a PTC disclosed herein comprises a mutation disclosed in Table 15.
  • the codon having the first sequence or the PTC comprises a mutation disclosed in Table 15.
  • the non-mutated, e.g., wildtype, codon sequence of the codon having the first sequence or the PTC is an original codon sequence provided in Table 15 and the amino acid corresponding to the non-mutated codon is an original AA provided in Table 15.
  • the TREM, TREM core fragment or TREM fragment recognizes a stop codon and mediates incorporation of the original AA provided in Table 15 at the position of the stop codon. In an embodiment, the TREM, TREM core fragment or TREM fragment recognizes a stop codon and mediates incorporation of an amino acid belonging to the same group as the original AA, e.g., as provided in Table 16. Other genetic abnormalities, such as insertions and/or deletions can also result in a PTC in an ORF. Table 15.
  • Nonpolar, aliphatic R group leucine Disclosed herein, inter alia, are endogenous ORFs comprising a codon having a first sequence, e.g., a mutation, e.g., a PTC.
  • An ORF having a PTC e.g., as described herein, can be present, or part of in any gene.
  • the ORF can be present or be part of any gene in the human genome.
  • a PTC disclosed herein is present in a gene disclosed in any one of Tables 17, 18, or 20.
  • Exemplary genes having ORFs comprising a PTC are provided in Table 3.
  • Table 17: Exemplary genes with ORFs having a PTC A 2ML1 ARFGEF1 CACNA1G CNOT1 DLG4 Attorney Docket No.: F2099-7037WO ACAD8 ASCC3 CARS2 COL9A1 DPH1 ACADL ASH1L CCDC140 COQ4 DPYD-AS1 Attorney Docket No.: F2099-7037WO FIBP GTPBP3 KCNMA1 MARS1 NCAPH2 FLAD1 HACE1 KCNQ5 MARS2 NCF2 Attorney Docket No.: F2099-7037WO P4HA1 PRDM12 RRM2B SLC18A3 SYN2 PAK6 PREPL RS1 SLC20A2 SYNJ1 Attorney Docket No.: F2099-7037WO T XNRD2 Z
  • the PTC is present within the FVIII gene and comprises an R mutation, e.g., an R2228X mutation.
  • the PTC is present within the FIX gene and comprises an R mutation, e.g., an R29X mutation, an R116X mutation, an R248X mutation, an R252X mutation, an R333X mutation, and/or an R338X mutation.
  • the PTC is present within the CFTR gene and comprises an R mutation, e.g., an R553X mutation.
  • the PTC is present within the MeCP2 gene and comprises an R mutation, e.g., an R168X mutation.
  • the PTC is present within the NAGLU gene and comprises an R mutation, e.g., an R626X mutation. In an embodiment, t the PTC is present within the DMD gene and comprises an R mutation, e.g., an R3881X mutation. In an embodiment, the PTC is present within the GAA gene and comprises an R mutation, e.g., an R854X mutation. In an embodiment, the PTC is present within the RP1 gene and comprises an R mutation, e.g., an R667X mutation. In an embodiment, the PTC is present within the RP2 gene and comprises an R mutation, e.g., an R120X mutation.
  • the PTC is present within the ABCA4 gene and comprises an R mutation, e.g., an R2030X mutation.
  • the PTC is present within the PCD gene and comprises an R mutation, e.g., an R245X mutation.
  • the PTC is present within the REP1 gene and comprises an R mutation, e.g., an R270X mutation.
  • the PTC is a mutation in the GLA gene, e.g., an R220X mutation and/or an R227X mutation.
  • the PTC is present within the MUT gene and comprises an R mutation, e.g., an R228X mutation, an R403X mutation, an R467X mutation, and/or an R727X mutation.
  • the PTC is present within the TP53 gene and comprises an R mutation, e.g., an R578X mutation.
  • the PTC is present within the ATM gene and comprises an R mutation, e.g., an R35X mutation.
  • Attorney Docket No.: F2099-7037WO Diseases or disorders associated with a PTC A TREM composition disclosed herein can be used treat a disorder or disease associated with a PTC, e.g., as described herein.
  • Exemplary diseases or disorders associated with a PTC are listed in Tables 18, 19, and 20.
  • the subject has a disease or disorder provided in any one of Tables 4- 6.
  • the cell is associated with, e.g., is obtained from a subject who has, a disorder or disease listed in any one of Tables 18-20.
  • the disorder or disease can be chosen from the left column of Table 18.
  • the disorder or disease is chosen from the left column of Table 18 and, in embodiments the PTC is in a gene chosen from the right column of Table 18, e.g., any one of the genes provided in the right column of Table 18.
  • the PTC is in a gene corresponding to the disorder or disease provided in the left column of Table 18.
  • the PTC can be at a position provided in Table 18.
  • the disorder or symptom is chosen from a disorder or disease provided in Table 19.
  • the disorder or symptom is chosen from a disorder or disease provided in Table 20.
  • the disorder or symptom is chosen from a disorder or disease provided in Table 20 and, in embodiments, the PTC is in any gene provided in Table 20.
  • the disorder or symptom is chosen from a disorder or disease provided in Table 20 and the PTC is in a corresponding gene provided in Table 20, e.g., a gene corresponding to the disease or disorder.
  • the disorder or symptom is chosen from a disorder or disease provided in Table 20 and the PTC is not in a gene provided in Table 20.
  • the PTC is at any position within the ORF of the gene, e.g., upstream of the naturally occurring stop codon.
  • Table 18 Exemplary diseases or disorders Disease/disorder or protein Exemplary Point Mutation
  • F2099-7037WO Lynch syndrome NM 000251.2(MSH2):c.212-1G>A Breast-ovarian cancer, familial 1 NM 007294.3(BRCA1):c.963G>A er
  • Attorney Docket No.: F2099-7037WO Angelman syndrome p.Trp768Ter
  • SMPD1 NM 000543.4(SMPD1):c.168G>A
  • p.Ile l l hr Attorney Docket No.: F2099-7037WO Table 19: Additional exemplary disorders 5q-syndrome Holt-Oram syndrome Adams-Oliver syndrome-1 Hypoparathyroidism d)
  • Attorney Docket No.: F2099-7037WO Table 20 Exemplary
  • the disease or disorder associated with a PTC is Fabry disease.
  • a TREM e.g., a TREM described herein
  • the level of a GLA protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, Attorney Docket No.: F2099-7037WO 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a GLA protein in a healthy, non-Fabry disease fibroblast).
  • a reference value e.g., level of a GLA protein in a healthy, non-Fabry disease fibroblast.
  • the disease or disorder associated with a PTC is a blood clotting disorder, e.g., Hemophilia B.
  • a TREM e.g., a TREM described herein
  • the level of a Factor IX (FIX) protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a FIX protein in a healthy, non-disease cell).
  • a reference value e.g., level of a FIX protein in a healthy, non-disease cell
  • the disease or disorder associated with a PTC is an autosomal recessive disorder, such as neuronal ceroid lipofuscinosis type 2 (CNL2).
  • a TREM e.g., a TREM described herein
  • the level of a tripeptidyl peptidase 1 (TPP1) protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a TPP1 protein in a healthy, non-disease cell).
  • a reference value e.g., level of a TPP1 protein in a healthy, non-disease cell
  • the disease or disorder associated with a PTC is a disease or disorder associated with hearing loss, such as Usher syndrome (e.g., Usher syndrome type 1F).
  • Usher syndrome e.g., Usher syndrome type 1F
  • the level of a protocadherin 15 precursor (PCDH15) protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a PCDH15 protein in a healthy, non-disease cell).
  • a reference value e.g., level of a PCDH15 protein in a healthy, non-disease cell
  • the disease or disorder associated with a PTC is a proliferative disease, such as a benign neoplasm or a cancer.
  • the proliferative disease is associated with a benign neoplasm.
  • a benign neoplasm may include adenoma, fibroma, hemangioma, tuberous sclerosis, and lipoma. All types of benign neoplasms disclosed herein or known in the art are contemplated as being within the scope of the disclosure.
  • the proliferative disease is a cancer.
  • cancer refers to a malignant neoplasm (Stedman’s Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990). All types of cancers disclosed herein or known in the art are contemplated as being within the scope of the disclosure.
  • Exemplary cancers include, but are not limited to, acoustic neuroma; adenocarcinoma; adrenal gland cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer; benign monoclonal gammopathy; biliary cancer (e.g., cholangiocarcinoma); bladder cancer; breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast); brain cancer (e.g., meningioma, glioblastomas, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma); bronchus cancer; carcinoid tumor; cervical cancer (e.g., cervical adenocarcinoma); choriocar
  • Wilms tumor, renal cell carcinoma); liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; myeloproliferative disorder (MPD) (e.g., polycythemia vera (PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.
  • HCC hepatocellular cancer
  • lung cancer e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung
  • myelofibrosis MF
  • chronic idiopathic myelofibrosis chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)
  • neuroblastoma e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis
  • neuroendocrine cancer e.g., gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor
  • osteosarcoma e.g., bone cancer
  • ovarian cancer e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma
  • papillary adenocarcinoma pancreatic cancer
  • pancreatic cancer e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), Islet cell tumors
  • the cancer is a solid tumor, such as a sarcoma or a carcinoma (e.g., lung cancer, brain cancer, breast cancer, bladder cancer, prostate cancer, colon cancer, rectal cancer).
  • a sarcoma or a carcinoma e.g., lung cancer, brain cancer, breast cancer, bladder cancer, prostate cancer, colon cancer, rectal cancer.
  • the present disclosure features methods of treating a disease or disorder in a cell or subject by administration of a TREM (e.g., a TREM described herein) to the cell or subject.
  • a TREM e.g., a TREM described herein
  • Exemplary diseases or disorders include hemophilias, aminoacidopathies, metal storage disorders, peroxisome biogenesis disorder, progressive rare lung disease, diseases related to lipid metabolism, diseases related to galactose metabolism, systemic organic acidemias, urea cycle disorders, cholestastis disorders, bilirubin metabolism disorders, lysososomal storage disorders, glycogen storage diseases, and oxalate metabolism disorders.
  • the disease or disorder is a hemophilia, e.g., hemophilia A or hemophilia B.
  • the disease or disorder is an aminoacidopathy, e.g., tyrosinemia type 1, tyrosinemia type 2, tyrosinemia type 3, maple syrup urine disease, alkaptonuria, or phenylketonuria.
  • the disease or disorder is a systemic organic acidemia, e.g., methylmalonic acidemia (MMUT), methylmalonic acidemia (non-MMUT), propionic acidemia type A, propionic acidemia type B, or isovaleric acidemia.
  • the disease or disorder is a urea cycle disorder, e.g, argininosuccinate lyase deficiency, argininosuccinate lyase deficiency-D, citrullinemia type 1, citrullinemia type 2, carbamoyl phosphate synthetase-D, ornithine transcarbamylase, arginemia, or hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome.
  • a urea cycle disorder e.g, argininosuccinate lyase deficiency, argininosuccinate lyase deficiency-D, citrullinemia type 1, citrullinemia type 2, carbamoyl phosphate synthetase-D, ornithine transcarbamylase, arginemia, or hyperornithinemia-hyperammonemia-homocitrullinuria (HHH)
  • the disease or disorder is lysosomal storage disorder, e.g., mucopolysaccharidosis 1, mucopolysaccharidosis 2, Fabry disease, lysosomal acid lipas deficiency, Pompe disease, Gaucher disease, Niemann Pick A, or Niemann Pick B.
  • the disease or disorder is a bilirubin metabolism disorder, e.g., Crigler-Najjar syndrome.
  • the disease or disorder is a cholestastis disorder, e.g., progressive familial intrahepatic cholestasis Attorney Docket No.: F2099-7037WO (PFIC) type 1, PFIC type 2, or PFIC type 3.
  • the disease or disorder is a disease related to lipid metabolism, e.g., sitosterolemia (ABCG5) or sitosterolemia (ABCG8).
  • the disease or disorder is a glycogen storage disease, e.g., glycogen storage disease 1a, glycogen storage disease 1b, or glycogen storage disease 3a.
  • the disease or disorder is a metal storage disorder, e.g., Wilson disease or hereditary hemochromatosis.
  • the disease or disorder is a progressive rare lung disease, e.g., alpha-1 antitrypsin deficiency.
  • the disease or disorder is a peroxisome biogenesis disorder, e.g., PBD RCDP1.
  • the disease or disorder is an oxalate metabolism disorder, e.g, primary hyperoxaluria type 1, primary hyperoxaluria type 2, or primary hyperoxaluria type 3.
  • the disease or disorder is a congenital disorder related to Notch signaling, e.g., Alagille syndrome.
  • the disease or disorder is an amyloidosis, e.g., familial amyloid polyneuropathy.
  • the present disclosure features a method of treating a disease or disorder in a subject, the method comprising administering to the subject a TREM comprising the nucleotide sequence of any one of the TREMS listed in FIG.1.
  • the disease or disorder is selected from a hemophilia, aminoacidopathy, metal storage disorder, peroxisome biogenesis disorder, progressive rare lung disease, disease related to lipid metabolism, disease related to galactose metabolism, systemic organic acidemia, urea cycle disorder, cholestastis disorder, bilirubin metabolism disorder, lysososomal storage disorder, glycogen storage disease, and oxalate metabolism disorder.
  • the TREM comprises the sequence of any one of SEQ ID NO: 622, 623, 624, 4249, 4386, 4834, 5630, 6707, 6749, 6947, or 8051, or a fragment or variant thereof.
  • TREMs, TREM core fragments, and TREM fragments In vitro methods for synthesizing oligonucleotides are known in the art and can be used to make a TREM, a TREM core fragment or a TREM fragment disclosed herein.
  • a TREM, TREM core fragment or TREM fragment can be synthesized using solid state synthesis or liquid phase synthesis.
  • a TREM, a TREM core fragment or a TREM fragment made according to an in vitro synthesis method disclosed herein has a different modification profile Attorney Docket No.: F2099-7037WO compared to a TREM expressed and isolated from a cell, or compared to a naturally occurring tRNA.
  • a TREM composition e.g., a TREM pharmaceutical composition
  • comprises a pharmaceutically acceptable excipient exemplary excipients include those provided in the FDA Inactive Ingredient Database (https://www.accessdata.fda.gov/scripts/cder/iig/index. Cfm).
  • a TREM composition e.g., a TREM pharmaceutical composition
  • a TREM composition e.g., a TREM pharmaceutical composition
  • a TREM composition e.g., a TREM pharmaceutical composition
  • a TREM composition comprises at least 1 x 10 6 TREM molecules, at least 1 x 10 7 TREM molecules, at least 1 x 10 8 TREM molecules or at least 1 x 10 9 TREM molecules. In an embodiment, a TREM composition comprises at least 1 x 10 6 TREM core fragment molecules, at least 1 x 10 7 TREM core fragment molecules, at least 1 x 10 8 TREM core fragment molecules or at least 1 x 10 9 TREM core fragment molecules. In an embodiment, a TREM composition comprises at least 1 x 10 6 TREM fragment molecules, at least 1 x 10 7 TREM fragment molecules, at least 1 x 10 8 TREM fragment molecules or at least 1 x 10 9 TREM fragment molecules.
  • a TREM composition produced by any of the methods of making disclosed herein can be charged with an amino acid using an in vitro charging reaction as known in the art.
  • a TREM composition comprise one or more species of TREMs, TREM core fragments, or TREM fragments.
  • a TREM composition comprises a single species of TREM, TREM core fragment, or TREM fragment.
  • a TREM composition comprises a first TREM, TREM core fragment, or TREM fragment species and a second TREM, TREM core fragment, or TREM fragment species.
  • the TREM, TREM core fragment, or TREM fragment has at least 70, 75, 80, 85, 90, or 95, or has 100%, identity with a sequence encoded by a nucleic acid in Table 1.
  • the TREM comprises a consensus sequence provided herein.
  • a TREM composition can be formulated as a liquid composition, as a lyophilized composition or as a frozen composition.
  • a TREM composition can be formulated to be suitable for pharmaceutical use, e.g., a pharmaceutical TREM composition.
  • a pharmaceutical TREM composition is substantially free of materials and/or reagents used to separate and/or purify a TREM, TREM core fragment, or TREM fragment.
  • a TREM composition can be formulated with water for injection.
  • a TREM composition formulated with water for injection is suitable for pharmaceutical use, e.g., comprises a pharmaceutical TREM composition.
  • TREM characterization A TREM, TREM core fragment, or TREM fragment, or a TREM composition, e.g., a pharmaceutical TREM composition, produced by any of the methods disclosed herein can be assessed for a characteristic associated with the TREM, TREM core fragment, or TREM fragment or the TREM composition, such as purity, sterility, concentration, structure, or functional activity of the TREM, TREM core fragment, or TREM fragment. Any of the above- mentioned characteristics can be evaluated by providing a value for the characteristic, e.g., by evaluating or testing the TREM, TREM core fragment, or TREM fragment, or the TREM Attorney Docket No.: F2099-7037WO composition, or an intermediate in the production of the TREM composition.
  • the value can also be compared with a standard or a reference value.
  • the TREM composition can be classified, e.g., as ready for release, meets production standard for human trials, complies with ISO standards, complies with cGMP standards, or complies with other pharmaceutical standards.
  • the TREM composition can be subjected to further processing, e.g., it can be divided into aliquots, e.g., into single or multi-dosage amounts, disposed in a container, e.g., an end-use vial, packaged, shipped, or put into commerce.
  • one or more of the characteristics can be modulated, processed or re-processed to optimize the TREM composition.
  • the TREM composition can be modulated, processed or re-processed to (i) increase the purity of the TREM composition; (ii) decrease the amount of fragments in the composition; (iii) decrease the amount of endotoxins in the composition; (iv) increase the in vitro translation activity of the composition; (v) increase the TREM concentration of the composition; or (vi) inactivate or remove any viral contaminants present in the composition, e.g., by reducing the pH of the composition or by filtration.
  • the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has a purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, i.e., by mass.
  • the TREM e.g., TREM composition or an intermediate in the production of the TREM composition
  • the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has low levels or absence of endotoxins, e.g., a negative result as measured by the Limulus amebocyte lysate (LAL) test.
  • the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has in-vitro translation activity, e.g., as measured by an assay described in Examples 6-7.
  • the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has a TREM Attorney Docket No.: F2099-7037WO concentration of at least 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10 ng/mL, 50 ng/mL, 0.1 ug/mL, 0.5 ug/mL,1 ug/mL, 2 ug/mL, 5 ug/mL, 10 ug/mL, 20 ug/mL, 30 ug/mL, 40 ug/mL, 50 ug/mL, 60 ug/mL, 70 ug/mL, 80 ug/mL, 100 ug/mL, 200 ug/mL, 300 ug/mL, 500 ug/mL, 1000 ug/mL, 5000 u
  • the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) is sterile, e.g., the composition or preparation supports the growth of fewer than 100 viable microorganisms as tested under aseptic conditions, the composition or preparation meets the standard of USP ⁇ 71>, and/or the composition or preparation meets the standard of USP ⁇ 85>.
  • the TREM, TREM core fragment, or TREM fragment e.g., TREM composition or an intermediate in the production of the TREM composition
  • any viral contaminant, e.g., residual virus, present in the composition is inactivated or removed.
  • any viral contaminant, e.g., residual virus is inactivated, e.g., by reducing the pH of the composition.
  • any viral contaminant, e.g., residual virus is removed, e.g., by filtration or other methods known in the field.
  • TREM administration Any TREM composition or pharmaceutical composition described herein can be administered to a cell, tissue or subject, e.g., by direct administration to a cell, tissue and/or an organ in vitro, ex-vivo or in vivo.
  • In-vivo administration may be via, e.g., by local, systemic and/or parenteral routes, for example intravenous, subcutaneous, intraperitoneal, intrathecal, intramuscular, ocular, nasal, urogenital, intradermal, dermal, enteral, intravitreal, intracerebral, intrathecal, or epidural.
  • Vectors and Carriers In some embodiments the TREM, TREM core fragment, or TREM fragment or TREM composition described herein, is delivered to cells, e.g. mammalian cells or human cells, using a vector.
  • the vector may be, e.g., a plasmid or a virus. In some embodiments, delivery is in vivo, in vitro, ex vivo, or in situ.
  • the virus is an adeno associated virus (AAV), a lentivirus, or an adenovirus.
  • AAV adeno associated virus
  • the system or components of the system are Attorney Docket No.: F2099-7037WO delivered to cells with a viral-like particle or a virosome.
  • the delivery uses more than one virus, viral-like particle or virosome.
  • Carriers A TREM, a TREM composition or a pharmaceutical TREM composition described herein may comprise, may be formulated with, or may be delivered in, a carrier.
  • the carrier may be a viral vector (e.g., a viral vector comprising a sequence encoding a TREM, a TREM core fragment or a TREM fragment).
  • the viral vector may be administered to a cell or to a subject (e.g., a human subject or animal model) to deliver a TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition.
  • a viral vector may be systemically or locally administered (e.g., injected).
  • Viral genomes provide a rich source of vectors that can be used for the efficient delivery of exogenous genes into a mammalian cell. Viral genomes are known in the art as useful vectors for delivery because the polynucleotides contained within such genomes are typically incorporated into the nuclear genome of a mammalian cell by generalized or specialized transduction.
  • viral vectors include a retrovirus (e.g., Retroviridae family viral vector), adenovirus (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvovirus (e.g., adeno-associated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e.g., influenza virus), rhabdovirus (e.g., rabies and vesicular stomatitis virus), paramyxovirus (e.g., measles and Sendai), positive strand RNA viruses, such as picornavirus and alphavirus, and double stranded DNA viruses including adenovirus, herpesvirus (e.g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus, replication deficient herpes virus), and poxvirus (e.g., Retroviridae family viral vector), adenovirus (e.g., Ad5, Ad26, Ad34
  • viruses include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, human papilloma virus, human foamy virus, and hepatitis virus, for example.
  • retroviruses include: avian leukosis-sarcoma, avian C-type viruses, mammalian C- type, B-type viruses, D-type viruses, oncoretroviruses, HTLV-BLV group, lentivirus, alpharetrovirus, gammaretrovirus, spumavirus (Coffin, J. M., Retroviridae: The viruses and their replication, Virology (Third Edition) Lippincott-Raven, Philadelphia, 1996).
  • murine leukemia viruses include murine leukemia viruses, murine sarcoma viruses, mouse mammary tumor virus, bovine Attorney Docket No.: F2099-7037WO leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, Gibbon ape leukemia virus, Mason Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus and lentiviruses.
  • vectors are described, for example, in US Patent No.5,801,030, the teachings of which are incorporated herein by reference.
  • the system or components of the system are delivered to cells with a viral-like particle or a virosome.
  • Cell and vesicle-based carriers A TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition described herein can be administered to a cell in a vesicle or other membrane-based carrier.
  • a TREM, a TREM core fragment or a TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein is administered in or via a cell, vesicle or other membrane-based carrier.
  • the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition can be formulated in liposomes or other similar vesicles.
  • Liposomes are spherical vesicle structures composed of a uni- or multilamellar lipid bilayer surrounding internal aqueous compartments and a relatively impermeable outer lipophilic phospholipid bilayer. Liposomes may be anionic, neutral or cationic.
  • Liposomes are biocompatible, nontoxic, can deliver both hydrophilic and lipophilic drug molecules, protect their cargo from degradation by plasma enzymes, and transport their load across biological membranes and the blood brain barrier (BBB) (see, e.g., Spuch and Navarro, Journal of Drug Delivery, vol.2011, Article ID 469679, 12 pages, 2011. doi:10.1155/2011/469679 for review).
  • Vesicles can be made from several different types of lipids; however, phospholipids are most commonly used to generate liposomes as drug carriers. Methods for preparation of multilamellar vesicle lipids are known in the art (see for example U.S. Pat.
  • vesicle formation can be spontaneous when a lipid film is mixed with an aqueous solution, it can also be expedited by applying force in the form of shaking by using a homogenizer, sonicator, or an extrusion apparatus (see, e.g., Spuch and Navarro, Journal of Drug Delivery, vol.2011, Article ID 469679, 12 pages, 2011. doi:10.1155/2011/469679 for review).
  • Extruded lipids can be prepared by extruding through filters of decreasing size, as described in Attorney Docket No.: F2099-7037WO Templeton et al., Nature Biotech, 15:647-652, 1997, the teachings of which relating to extruded lipid preparation are incorporated herein by reference.
  • Lipid Nanoformulations/Lipid-based carriers Lipid nanoparticles are another example of a carrier that provides a biocompatible and biodegradable delivery system for the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition described herein.
  • Nanostructured lipid carriers are modified solid lipid nanoparticles (SLNs) that retain the characteristics of the SLN, improve drug stability and loading capacity, and prevent drug leakage.
  • Polymer nanoparticles are an important component of drug delivery. These nanoparticles can effectively direct drug delivery to specific targets and improve drug stability and controlled drug release.
  • Lipid–polymer nanoparticles a new type of carrier that combines liposomes and polymers, may also be employed. These nanoparticles possess the complementary advantages of PNPs and liposomes.
  • a PLN is composed of a core–shell structure; the polymer core provides a stable structure, and the phospholipid shell offers good biocompatibility.
  • lipid nanoparticles are disclosed in International Application PCT/US2014/053907, the entire contents of which are hereby incorporated by reference.
  • an LNP described in paragraphs [403-406] or [410-413] of PCT/US2014/053907 can be used as a carrier for the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition described herein.
  • Additional exemplary lipid nanoparticles are disclosed in U.S.
  • Patent 10,562,849 the entire contents of which are hereby incorporated by reference.
  • an LNP of formula (I) as described in columns 1-3 of U.S. Patent 10,562,849 can be used as a carrier for the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition described herein.
  • Lipids that can be used in nanoparticle formations include, for example those described in Table 4 of WO2019217941, which is incorporated by reference, e.g., a lipid-containing nanoparticle can comprise one or more of the lipids in Table 4 of Attorney Docket No.: F2099-7037WO WO2019217941.
  • Lipid nanoparticles can include additional elements, such as polymers, such as the polymers described in Table 5 of WO2019217941, incorporated by reference.
  • 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 dialkoxyprop
  • sterols that can be incorporated into lipid nanoparticles include one or more of cholesterol or cholesterol derivatives, such as those in W02009/127060 or US2010/0130588, which are incorporated by reference. Additional exemplary sterols include phytosterols, including those described in Eygeris et al (2020), incorporated herein by reference.
  • Cationic Lipids (Positively Charged) and Ionizable Lipids 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.
  • the cationic lipid is a lipid capable of being positively charged, e.g., under physiological conditions.
  • exemplary cationic lipids include one or more amine group(s) which bear the positive charge.
  • 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), 3 ⁇ -[N-(N',N'- dimethylaminoethyl)carbamoyl) cholesterol (DC-chol), 1,2-dioleoyloxy-3-[trimethylammonio]- propane (DOTAP), 1,2-dioctadecyloxy-3-[trimethylammonio]-propane (DSTAP), and 1,2- dioleoyloxypropyl-3-
  • the lipid-based carrier comprises more than one cationic lipid.
  • the lipid-based carrier comprises a cationic lipid having an effective pKa over 6.0.
  • the lipid-based carrier further comprises a second cationic lipid having a different effective pKa (e.g., greater than the first effective pKa) than the first cationic lipid.
  • 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, which is incorporated by reference.
  • the cationic lipid is an ionizable lipid (e.g., a lipid that is protonated at low pH, but that remains neutral at physiological pH).
  • 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, , , Attorney Docket No.: F2099-7037WO , entirety).
  • 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), which is incorporated herein by reference in its entirety.
  • 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), (which is incorporated by reference herein in its entirety).
  • 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; I, IA, 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;
  • the lipid-based carrier (or lipid nanoformulation) further includes biodegradable ionizable lipids, for instance, (9Z,l2Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,l2-dienoate, also called 3- ((4,4- bis(octyloxy)butanoyl)oxy)-2-(((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,l2Z)-octadeca-9,l2-dienoate).
  • biodegradable ionizable lipids for instance, (9Z,l2Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbon
  • the lipid particle comprises an ionizable lipid, a non-cationic lipid, a conjugated lipid that inhibits aggregation of particles, and a sterol.
  • the amounts of these components can be varied independently and to achieve desired properties.
  • the lipid nanoparticle comprises an ionizable lipid is in an amount from about 20 mol % to about 90 mol % of the total lipids (in other embodiments it may be 20-70% (mol), 30-60% (mol) or 40-50% (mol); about 50 mol % to about 90 mol % of the total lipid Attorney Docket No.: F2099-7037WO present in the lipid nanoparticle), a non-cationic lipid in an amount from about 5 mol % to about 30 mol % of the total lipids, a conjugated lipid in an amount from about 0.5 mol % to about 20 mol % of the total lipids, and a sterol in an amount from about 20 mol % to about 50 mol % of the total lipids.
  • the ratio of total lipid to nucleic acid can be varied as desired.
  • the total lipid to nucleic acid (mass or weight) ratio can be from about 10: 1 to about 30: 1.
  • the lipid to nucleic acid 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 lipids and nucleic acid can be adjusted to provide a desired N/P ratio, for example, N/P ratio of 3, 4, 5, 6, 7, 8, 9, 10 or higher.
  • the lipid nanoparticle formulation s overall lipid content can range from about 5 mg/ml to about 30 mg/mL.
  • Some non-limiting example of lipid compounds that may be used (e.g., in combination with other lipid components) to form lipid nanoparticles for the delivery of compositions described herein, e.g., nucleic acid (e.g., RNA) described herein includes, (i) a TREM composition described herein to the liver and/or hepatocyte cells. a TREM composition described herein to the liver and/or hepatocyte cells.
  • an LNP comprising Formula (iii) is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells. to deliver a TREM composition described herein to the liver and/or hepatocyte cells. is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells.
  • an LNP comprising Formula (ix) is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells.
  • an LNP comprising Formula (xii) is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells. TREM composition described herein to the liver and/or hepatocyte cells.
  • an LNP comprises a compound of Formula (xiii) and a compound of Formula (xiv).
  • Attorney Docket No.: F2099-7037WO a TREM an LNP comprising a formulation of Formula (xvi) is used to deliver a TREM composition described herein to the lung endothelial cells.
  • lipid nanoparticles for the delivery of e.g., a herein is made by one of the following reactions: (a) provided in an LNP that comprises an ionizable lipid.
  • the ionizable lipid is heptadecan-9-yl 8-((2-hydroxyethyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate (SM-102); e.g., as described in Example 1 of US9,867,888 (incorporated by reference herein in its entirety).
  • the ionizable lipid is 9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,12-dienoate (LP01), e.g., as synthesized in Example 13 of WO2015/095340 (incorporated by reference herein in its entirety).
  • the ionizable lipid is Di((Z)-non-2-en-1-yl) 9-((4-dimethylamino)- butanoyl)oxy)heptadecanedioate (L319), e.g. as synthesized in Example 7, 8, or 9 of US2012/0027803 (incorporated by reference herein in its entirety).
  • the ionizable lipid is 1,1’-((2-(4-(2-((2-(Bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl) amino)ethyl)piperazin-1-yl)ethyl)azanediyl)bis(dodecan-2-ol) (C12-200), e.g., as synthesized in Examples 14 and 16 of WO2010/053572 (incorporated by reference herein in its entirety).
  • the ionizable lipid is Imidazole cholesterol ester (ICE) lipid (3S, 10R, 13R, 17R)-10, 13-dimethyl-17- ((R)-6-methylheptan-2-yl)-2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-lH- cyclopenta[a]phenanthren-3-yl 3-(1H-imidazol-4-yl)propanoate, e.g., Structure (I) from WO2020/106946 (incorporated by reference herein in its entirety).
  • ICE Imidazole cholesterol ester
  • an ionizable lipid may be a cationic lipid, an ionizable cationic lipid, 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.
  • the cationic lipid is a lipid capable of being positively charged, e.g., under physiological conditions.
  • Exemplary cationic lipids include one or more amine group(s) which bear the positive charge.
  • the lipid particle comprises a cationic lipid in formulation with one or more of neutral lipids, ionizable amine-containing lipids, biodegradable alkyne lipids, steroids, phospholipids including polyunsaturated lipids, structural lipids (e.g., sterols), PEG, cholesterol and polymer conjugated lipids.
  • the cationic lipid may be an ionizable cationic lipid.
  • An exemplary cationic lipid as disclosed herein may have an effective pKa over 6.0.
  • a lipid nanoparticle may comprise a second cationic lipid having a different effective pKa (e.g., greater than the first effective pKa), than the first cationic lipid.
  • a lipid nanoparticle may comprise between 40 and 60 mol percent of a cationic lipid, a neutral lipid, a steroid, a polymer conjugated lipid, and a therapeutic agent, e.g., a TREM described herein, encapsulated within or associated with the lipid nanoparticle.
  • the TREM is co-formulated with the cationic lipid.
  • the TREM may be adsorbed to the surface of an LNP, e.g., an LNP comprising a cationic lipid.
  • the TREM may be encapsulated in an LNP, e.g., an LNP comprising a cationic lipid.
  • the lipid nanoparticle may comprise a targeting moiety, e.g., coated with a targeting agent.
  • the LNP formulation is biodegradable.
  • a lipid nanoparticle comprising one or more lipid described herein, e.g., Formula (i), (ii), (ii), (vii) and/or (ix) encapsulates at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 92%, at least 95%, at least 97%, at least 98% or 100% of a TREM.
  • Exemplary ionizable lipids that can be used in lipid nanoparticle formulations include, without limitation, those listed in Table 1 of WO2019051289, incorporated herein by reference.
  • Additional exemplary lipids include, without limitation, one or more of the following formulae: X of US2016/0311759; I of US20150376115 or in US2016/0376224; I, II or III of US20160151284; I, IA, II, or IIA of US20170210967; I-c of US20150140070; A of US2013/0178541; I of US2013/0303587 or US2013/0123338; I of US2015/0141678; II, III, IV, or V of US2015/0239926; I of US2017/0119904; I or II of WO2017/117528; A of Attorney Docket No.: F2099-7037WO US2012/0149894; A of US2015/0057373; A of WO2013/116126; A of US2013/0090372; A of US2013/0274523; A of US2013/0274504; A of US2013/0053572; A of W02013/016058; A of
  • the ionizable lipid is MC3 (6Z,9Z,28Z,3 lZ)-heptatriaconta- 6,9,28,3 l-tetraen-l9-yl-4-(dimethylamino) butanoate (DLin-MC3-DMA or MC3), e.g., as described in Example 9 of WO2019051289A9 (incorporated by reference herein in its entirety).
  • the ionizable lipid is the lipid ATX-002, e.g., as described in Example 10 of WO2019051289A9 (incorporated by reference herein in its entirety).
  • the ionizable lipid is (l3Z,l6Z)-A,A-dimethyl-3- nonyldocosa-l3, l6-dien-l-amine (Compound 32), e.g., as described in Example 11 of WO2019051289A9 (incorporated by reference herein in its entirety).
  • the ionizable lipid is Compound 6 or Compound 22, e.g., as described in Example 12 of WO2019051289A9 (incorporated by reference herein in its entirety).
  • Non-Cationic Lipids e.g., Phospholipids
  • the lipid-based carrier or lipid nanoformulation
  • the non-cationic lipid is a phospholipid. In some embodiments, the non-cationic lipid is a phospholipid substitute or Attorney Docket No.: F2099-7037WO replacement. In some embodiments, the non-cationic lipid is a negatively charged (anionic) lipid.
  • 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 (
  • acyl groups in these lipids are preferably acyl groups derived from fatty acids having C 10- C 24 carbon chains, e.g., lauroyl, myristoyl, paimitoyl, stearoyl, or oleoyl.
  • Additional exemplary lipids include, without limitation, those described in Kim et al. (2020) dx.doi.org/10.1021/acs.nanolett.0c01386, which is incorporated herein by reference.
  • lipids include, in some embodiments, plant lipids found to improve liver transfection with mRNA (e.g., DGTS).
  • the lipid-based carrier (or lipid nanoformulation) may comprise a combination of distearoylphosphatidylcholine/cholesterol, Attorney Docket No.: F2099-7037WO dipalmitoylphosphatidylcholine/cholesterol, dimyrystoylphosphatidylcholine/cholesterol, 1,2- Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/cholesterol, or egg sphingomyelin/cholesterol.
  • DOPC 1,2- Dioleoyl-sn-glycero-3-phosphocholine
  • 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.
  • nonphosphorous lipids such as, e.g., stearylamine, dodecylamine, hexadecylamine, acetyl palmitate, glycerol ricinoleate, hexadecyl stearate, isopropyl my
  • 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, which is incorporated herein by reference in its entirety.
  • 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.
  • 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).
  • the lipid-based carrier (or lipid nanoformulation) does not include any phospholipids.
  • 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).
  • 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
  • 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 include, without limitation, those described in Kim et al. (2020) dx.doi.org/10.1021/acs.nanolett.0c01386, incorporated herein by reference.
  • Such lipids include, in some embodiments, plant lipids found to improve liver transfection with mRNA (e.g., DGTS).
  • Other examples of non-cationic lipids suitable for use in the lipid nanoparticles include, without limitation, nonphosphorous lipids such as, e.g., stearylamine, dodeeylamine, hexadecylamine, acetyl palmitate, glycerol ricinoleate, hexadecyl stereate, isopropyl myristate, amphoteric acrylic polymers, triethanolamine-lauryl sulfate, alkyl-aryl sulfate polyethyloxylated fatty acid amides, dioctadecyl dimethyl ammonium bromide, ceramide, sphingomyelin, and the like.
  • non-cationic lipids are described in WO2017/099823 or US patent publication US2018/0028664, the contents of which is incorporated herein by reference in their entirety.
  • the non-cationic lipid is oleic acid or a compound of Formula I, II, or IV of US2018/0028664, incorporated herein by reference in its entirety.
  • the non-cationic lipid can comprise, for example, 0-30% (mol) of the total lipid present in the lipid nanoparticle. In some embodiments, the non-cationic lipid content is 5-20% (mol) or 10-15% (mol) of the total lipid present in the lipid nanoparticle.
  • the molar ratio of ionizable lipid to the 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).
  • Attorney Docket No.: F2099-7037WO the lipid nanoparticles do not comprise any phospholipids.
  • Structural Lipids The lipid-based carrier (or lipid nanoformulation) described herein may further comprise one or more structural lipids.
  • structural lipid refers to sterols (e.g., cholesterol) and also to lipids containing sterol moieties.
  • 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.
  • the structural lipid is a sterol.
  • the structural lipid is a steroid.
  • the structural lipid is cholesterol.
  • the structural lipid is an analog of cholesterol.
  • the structural lipid is alpha-tocopherol.
  • structural lipids may be incorporated into the lipid-based carrier at molar ratios ranging from about 0.1 to 1.0 (cholesterol phospholipid).
  • 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, which are incorporated herein by reference in their entirety. Additional exemplary sterols include phytosterols, including those described in Eygeris et al. (2020), Nano Lett.2020;20(6):4543- 4549, incorporated herein by reference.
  • 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.
  • the cholesterol derivative is a polar analogue, e.g., cholesteryl-(4'-hydroxy)-buty1 ether.
  • 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.
  • a component such as a sterol, to provide membrane integrity.
  • sterol that can be used in the lipid nanoparticle is cholesterol and derivatives thereof.
  • cholesterol derivatives include polar analogues such as 5a-choiestanol, 53-coprostanol, choiesteryl-(2 , - hydroxy)-ethyl ether, choiesteryl-(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.
  • polar analogues such as 5a-choiestanol, 53-coprostanol, choiesteryl-(2 , - hydroxy)-ethyl ether, choiesteryl-(4’- hydroxy)-butyl ether, and 6-ketocholestanol
  • non-polar analogues such as 5a-
  • the cholesterol derivative is a polar analogue, e.g., choiesteryl-(4 ‘-hydroxy)-buty1 ether.
  • exemplary cholesterol derivatives are described in PCT publication W02009/127060 and US patent publication US2010/0130588, each of which is incorporated herein by reference in its entirety.
  • the component providing membrane integrity such as a sterol, can comprise 0-50% (mol) (e.g., 0-10%, 10-20%, 20-30%, 30-40%, or 40-50%) of the total lipid present in the lipid nanoparticle. In some embodiments, such a component is 20-50% (mol) 30- 40% (mol) of the total lipid content of the lipid nanoparticle.
  • the lipid-based carrier may include one or more polymers or co-polymers, e.g., poly(lactic-co-glycolic acid) (PFAG) nanoparticles.
  • the lipid-based carrier may include one or more polyethylene glycol (PEG) lipid.
  • PEG-lipids examples include, but are not limited to, 1,2-Diacyl-sn-Glycero-3- Phosphoethanolamine-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); 1,2-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
  • the PEG lipid is a polyethyleneglycol-diacylglycerol (i.e., polyethyleneglycol diacylglycerol (PEG-DAG), PEG-cholesterol, or PEG-DMB) conjugate.
  • 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, which is incorporated herein by reference in its entirety).
  • 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).
  • one or more ionic lipids e.g., non-cationic lipid such as a neutral or anionic, or zwitterionic lipid
  • one or more sterols e.g., cholesterol
  • 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 (C18).
  • a PEG-dilaurylglycerol C12
  • PEG- dimyristylglycerol C14
  • PEG-dipalmitoylglycerol C16
  • PEG-disterylglycamide C18
  • 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-
  • DAG P
  • 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, all of which are incorporated herein by reference in their entirety.
  • 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.
  • the PEG-lipid is of Formula II of US 2015/0376115 or US 2016/0376224, both of which are incorporated herein by reference in their entirety.
  • the PEG-DAA conjugate can be, for example, PEG-dilauryloxypropyl, PEG- dimyristyloxypropyl, PEG-dipalmityloxypropyl, or PEG-distearyloxypropyl.
  • the PEG-lipid Attorney Docket No.: F2099-7037WO includes one of the following: used in place of PEG-lipid.
  • 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.
  • 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, which is incorporated herein by reference in its entirety.
  • the conjugated lipid e.g., the PEGylated lipid
  • the conjugated 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).
  • the conjugated lipid (e.g., the PEGylated lipid) content is 0.5-10 mol% or 2-5 mol% of the total lipid components.
  • 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).
  • 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.
  • the lipid nanoparticle can comprise a polyethylene glycol (PEG) or a conjugated lipid molecule. Generally, these are used to inhibit aggregation of lipid nanoparticles and/or provide steric stabilization.
  • PEG polyethylene glycol
  • conjugated lipids include, but are not limited to, PEG-lipid conjugates, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (such as ATTA-lipid conjugates), cationic-polymer lipid (CPL) conjugates, and mixtures thereof.
  • the conjugated lipid molecule is a PEG-lipid conjugate, for example, a (methoxy polyethylene glycol)-conjugated lipid.
  • PEG-lipid conjugates include, but are not limited to, 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)-l,2- distearoyl-sn-glycero-3-
  • exemplary PEG-lipid conjugates are described, for example, in US5,885,6l3, US6,287,59l, US2003/0077829, US2003/0077829, US2005/0175682, US2008/0020058, US2011/0117125, US2010/0130588, US2016/0376224, US2017/0119904, and US/099823, the contents of all of which are incorporated herein by reference in their entirety.
  • a PEG-lipid is a compound of Formula III, III-a-I, III-a-2, III-b-1, III-b-2, or V of US2018/0028664, the content of which is incorporated herein by reference in its entirety.
  • a PEG-lipid is of Formula II of US20150376115 or US2016/0376224, the content of both of which is incorporated herein by reference in its entirety.
  • the PEG-DAA conjugate can be, for example, PEG-dilauryloxypropyl, PEG- dimyristyloxypropyl, PEG- dipalmityloxypropyl, or PEG-distearyloxypropyl.
  • the PEG-lipid can be one or more of PEG- DMG, PEG-dilaurylglycerol, PEG-dipalmitoylglycerol, PEG- disterylglycerol, PEG- dilaurylglycamide, PEG-dimyristylglycamide, PEG- dipalmitoylglycamide, PEG- disterylglycamide, PEG-cholesterol (l-[8’-(Cholest-5-en-3[beta]- oxy)carboxamido-3’,6’- dioxaoctanyl] carbamoyl-[omega]-methyl-poly(ethylene glycol), PEG- DMB (3,4- Ditetradecoxylbenzyl- [omega]-methyl-poly(ethylene glycol) ether), and 1,2- dimyristoyl-sn- glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-
  • the PEG-lipid comprises PEG-DMG, 1,2- dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000].
  • the PEG-lipid comprises a structure selected from: can also be used in place of PEG-lipid.
  • 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.
  • conjugated lipids i.e., PEG-lipids, (POZ)-lipid conjugates, ATTA-lipid conjugates and cationic polymer-lipids are described in the PCT and LIS patent applications listed in Table 2 of WO2019051289A9, the contents of all of which are incorporated herein by reference in their entirety.
  • the PEG or the conjugated lipid can comprise 0-20% (mol) of the total lipid present in the lipid nanoparticle.
  • PEG or the conjugated lipid content is 0.5- 10% or 2-5% (mol) of the total lipid present in the lipid nanoparticle.
  • the lipid particle can comprise 30-70% ionizable lipid by mole or by total weight of the composition, 0-60% cholesterol by mole or by total weight of the composition, 0- 30% non-cationic-lipid by mole or by total weight of the composition and 1-10% conjugated lipid by mole or by total weight of the composition.
  • the composition comprises 30- 40% ionizable lipid by mole or by total weight of the composition, 40-50% cholesterol by mole Attorney Docket No.: F2099-7037WO or by total weight of the composition, and 10- 20% non-cationic-lipid by mole or by total weight of the composition.
  • the composition is 50-75% ionizable lipid by mole or by total weight of the composition, 20-40% cholesterol by mole or by total weight of the composition, and 5 to 10% non-cationic-lipid, by mole or by total weight of the composition and 1-10% conjugated lipid by mole or by total weight of the composition.
  • the composition may contain 60-70% ionizable lipid by mole or by total weight of the composition, 25-35% cholesterol by mole or by total weight of the composition, and 5-10% non-cationic-lipid by mole or by total weight of the composition.
  • the composition may also contain up to 90% ionizable lipid by mole or by total weight of the composition and 2 to 15% non-cationic lipid by mole or by total weight of the composition.
  • the formulation may also be a lipid nanoparticle formulation, for example comprising 8-30% ionizable lipid by mole or by total weight of the composition, 5- 30% non- cationic lipid by mole or by total weight of the composition, and 0-20% cholesterol by mole or by total weight of the composition; 4-25% ionizable lipid by mole or by total weight of the composition, 4-25% non-cationic lipid by mole or by total weight of the composition, 2 to 25% cholesterol by mole or by total weight of the composition, 10 to 35% conjugate lipid by mole or by total weight of the composition, and 5% cholesterol by mole or by total weight of the composition; or 2-30% ionizable lipid by mole or by total weight of the composition, 2-30% non-cationic lipid by mole or by total weight of the composition, 1 to 15% cholesterol by mole or by total weight of the composition, 2 to 35% conjugate lipid by mole or by total weight of the composition, and 1-20% cholesterol by mole or by total weight of the
  • the lipid particle formulation comprises ionizable lipid, phospholipid, cholesterol and a PEG-ylated lipid in a molar ratio of 50: 10:38.5: 1.5. In some other embodiments, the lipid particle formulation comprises ionizable lipid, cholesterol and a PEG-ylated lipid in a molar ratio of 60:38.5: 1.5. In some embodiments, the lipid particle comprises ionizable lipid, non-cationic lipid (e.g.
  • phospholipid e.g., cholesterol
  • sterol e.g., cholesterol
  • PEG-ylated lipid where the molar ratio of lipids ranges from 20 to 70 mole percent for the ionizable lipid, with a target of 40-60, the mole percent of non-cationic lipid ranges from 0 to 30, with a target of 0 to 15, the mole percent of sterol Attorney Docket No.: F2099-7037WO ranges from 20 to 70, with a target of 30 to 50, and the mole percent of PEG-ylated lipid ranges from 1 to 6, with a target of 2 to 5.
  • LNPs are directed to specific tissues by the addition of targeting domains.
  • biological ligands may be displayed on the surface of LNPs to enhance interaction with cells displaying cognate receptors, thus driving association with and cargo delivery to tissues wherein cells express the receptor.
  • the biological ligand may be a ligand that drives delivery to the liver, e.g., LNPs that display GalNAc result in delivery of nucleic acid cargo to hepatocytes that display asialoglycoprotein receptor (ASGPR).
  • ASGPR asialoglycoprotein receptor
  • the work of Akinc et al. Mol Ther 18(7):1357-1364 (2010) teaches the conjugation of a trivalent GalNAc ligand to a PEG-lipid (GalNAc-PEG-DSG) to yield LNPs dependent on ASGPR for observable LNP cargo effect (see, e.g., FIG.6 of Akinc et al.2010, supra).
  • ligand- displaying LNP formulations e.g., incorporating folate, transferrin, or antibodies
  • WO2017223135 which is incorporated herein by reference in its entirety, in addition to the references used therein, namely Kolhatkar et al., Curr Drug Discov Technol.20118:197-206; Musacchio and Torchilin, Front Biosci.201116:1388-1412; Yu et al., Mol Membr Biol.2010 27:286-298; Patil et al., Crit Rev Ther Drug Carrier Syst.200825:1-61 ; Benoit et al., Biomacromolecules.201112:2708-2714; Zhao et al., Expert Opin Drug Deliv.20085:309-319; Akinc et al., Mol Ther.201018:1357-1364; Srinivasan et al., Methods Mol Biol.2012820:105- Attorney Docket No.:
  • LNPs are selected for tissue-specific activity by the addition of a Selective ORgan Targeting (SORT) molecule to a formulation comprising traditional components, such as ionizable cationic lipids, amphipathic phospholipids, cholesterol and poly(ethylene glycol) (PEG) lipids.
  • SORT Selective ORgan Targeting
  • Nat Nanotechnol 15(4):313- 320 demonstrate that the addition of a supplemental “SORT” component precisely alters the in vivo RNA delivery profile and mediates tissue-specific (e.g., lungs, liver, spleen) gene delivery and editing as a function of the percentage and biophysical property of the SORT molecule.
  • the LNPs comprise biodegradable, ionizable lipids.
  • the LNPs comprise (9Z,l2Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,l2-dienoate, also called 3- ((4,4- bis(octyloxy)butanoyl)oxy)-2-(((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,l2Z)-octadeca-9,l2-dienoate) or another ionizable lipid.
  • the term cationic and ionizable in the context of LNP lipids is interchangeable, e.g., wherein ionizable lipids are cationic depending on the pH.
  • the average LNP diameter of the LNP formulation may be between 10s of nm and 100s of nm, e.g., measured by dynamic light scattering (DLS).
  • the average LNP diameter of the LNP formulation may be from about 40 nm to about 150 nm, such as about 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm.
  • the average LNP diameter of the LNP formulation may be from about 50 nm to about 100 nm, from about 50 nm to about 90 nm, from about 50 nm to about 80 nm, from about 50 nm to about 70 nm, from about 50 nm to about 60 nm, from about 60 nm to about 100 nm, from about 60 nm to about 90 nm, from about 60 nm to about 80 nm, from about 60 nm to about 70 nm, from about 70 nm to about 100 nm, from about Attorney Docket No.: F2099-7037WO 70 nm to about 90 nm, from about 70 nm to about 80 nm, from about 80 nm to about 100 nm, from about 80 nm to about 90 nm, or from about 90 nm to about 100 nm.
  • the average LNP diameter of the LNP formulation may be from about 70 nm to about 100 nm. In a particular embodiment, the average LNP diameter of the LNP formulation may be about 80 nm. In some embodiments, the average LNP diameter of the LNP formulation may be about 100 nm. In some embodiments, the average LNP diameter of the LNP formulation ranges from about l mm to about 500 mm, from about 5 mm to about 200 mm, from about 10 mm to about 100 mm, from about 20 mm to about 80 mm, from about 25 mm to about 60 mm, from about 30 mm to about 55 mm, from about 35 mm to about 50 mm, or from about 38 mm to about 42 mm.
  • a LNP may, in some instances, be relatively homogenous.
  • a polydispersity index may be used to indicate the homogeneity of a LNP, e.g., the particle size distribution of the lipid nanoparticles.
  • a small (e.g., less than 0.3) polydispersity index generally indicates a narrow particle size distribution.
  • a LNP may have a polydispersity index from about 0 to about 0.25, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, or 0.25.
  • the polydispersity index of a LNP may be from about 0.10 to about 0.20.
  • the zeta potential of a LNP may be used to indicate the electrokinetic potential of the composition.
  • the zeta potential may describe the surface charge of an LNP. Lipid nanoparticles with relatively low charges, positive or negative, are generally desirable, as more highly charged species may interact undesirably with cells, tissues, and other elements in the body.
  • the zeta potential of a LNP may be from about -10 mV to about +20 mV, from about -10 mV to about +15 mV, from about -10 mV to about +10 mV, from about -10 mV to about +5 mV, from about -10 mV to about 0 mV, from about -10 mV to about -5 mV, from about -5 mV to about +20 mV, from about -5 mV to about +15 mV, from about -5 mV to about +10 mV, from about -5 mV to about +5 mV, from about -5 mV to about 0 mV, from about 0 mV to about +20 mV, from about 0 mV to about +15 mV, from about 0 mV to about +10 mV, from about 0 mV to about +5 mV, from about 0 mV to about +20 mV, from
  • the efficiency of encapsulation of a TREM describes the amount of TREM that is encapsulated or otherwise associated with a LNP after preparation, relative to the initial amount provided.
  • the encapsulation efficiency is desirably high (e.g., close to 100%).
  • the encapsulation Attorney Docket No.: F2099-7037WO efficiency may be measured, for example, by comparing the amount of TREM in a solution containing the lipid nanoparticle before and after breaking up the lipid nanoparticle with one or more organic solvents or detergents.
  • An anion exchange resin may be used to measure the amount of free protein or nucleic acid (e.g., RNA) in a solution. Fluorescence may be used to measure the amount of free TREM in a solution.
  • the encapsulation efficiency of a TREM 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 80%. In some embodiments, the encapsulation efficiency may be at least 90%. In some embodiments, the encapsulation efficiency may be at least 95%.
  • a LNP may optionally comprise one or more coatings. In some embodiments, a LNP may be formulated in a capsule, film, or table having a coating.
  • a capsule, film, or tablet including a composition described herein may have any useful size, tensile strength, hardness or density. Additional exemplary lipids, formulations, methods, and characterization of LNPs are taught by WO2020061457, which is incorporated herein by reference in its entirety.
  • in vitro or ex vivo cell lipofections are performed using Lipofectamine MessengerMax (Thermo Fisher) or TransIT-mRNA Transfection Reagent (Mirus Bio).
  • LNPs are formulated using the GenVoy_ILM ionizable lipid mix (Precision NanoSystems).
  • LNPs are formulated using 2,2 ⁇ dilinoleyl ⁇ 4 ⁇ dimethylaminoethyl ⁇ [1,3] ⁇ dioxolane (DLin ⁇ KC2 ⁇ DMA) or dilinoleylmethyl ⁇ 4 ⁇ dimethylaminobutyrate (DLin-MC3-DMA or MC3), the formulation and in vivo use of which are taught in Jayaraman et al. Angew Chem Int Ed Engl 51(34):8529-8533 (2012), incorporated herein by reference in its entirety.
  • DLin ⁇ KC2 ⁇ DMA 2,2 ⁇ dilinoleyl ⁇ 4 ⁇ dimethylaminoethyl ⁇ [1,3] ⁇ dioxolane
  • DLin-MC3-DMA or MC3 dilinoleylmethyl ⁇ 4 ⁇ dimethylaminobutyrate
  • LNP formulations optimized for the delivery of CRISPR-Cas systems e.g., Cas9-gRNA RNP, gRNA, Cas9 mRNA
  • Cas9-gRNA RNP gRNA
  • Cas9 mRNA gRNA
  • Additional specific LNP formulations useful for delivery of nucleic acids are described in US8158601 and US8168775, both incorporated by reference, which include formulations used in patisiran, sold under the name ONPATTRO.
  • Attorney Docket No.: F2099-7037WO Exosomes can also be used as drug delivery vehicles for the TREM, TREM core fragment, TREM fragment, or TREM compositions or pharmaceutical TREM composition described herein.
  • TREM TREM core fragment
  • TREM fragment gRNA
  • TREM compositions pharmaceutical TREM composition described herein.
  • Ex vivo differentiated red blood cells can also be used as a carrier for a TREM, TREM core fragment, TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein. See, e.g., WO2015073587; WO2017123646; WO2017123644; WO2018102740; wO2016183482; WO2015153102; WO2018151829; WO2018009838; Shi et al.2014.
  • Fusosome compositions can also be used as carriers to deliver the TREM, TREM core fragment, TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein.
  • Virosomes and virus-like particles can also be used as carriers to deliver a TREM, TREM core fragment, TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein to targeted cells.
  • the lipid-based carrier (or lipid nanoformulation) comprises one of more of the compounds described herein, 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.
  • a non-cationic lipid e.g., a phospholipid
  • a sterol e.g., a sterol
  • a neutral lipid e.g., a neutral lipid
  • optionally conjugated lipid e.g., a PEGylated lipid
  • the lipid-based carrier further comprises a TREM (e.g., a TREM described herein).
  • TREM e.g., a TREM described herein.
  • 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 Attorney Docket No.: F2099-7037WO 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%,
  • a non-cationic lipid e.g.
  • 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.
  • the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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.
  • the encapsulation efficiency of the payload may be at least 70%.
  • 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.
  • the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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.
  • the encapsulation efficiency of the payload may be at least 70%.
  • 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 Attorney Docket No.: F2099-7037WO mol% (e.g., 1-5 mol%, or 1.5-2.5 mol%) PEGylated lipid.
  • the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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.
  • TREM e.g., a TREM described herein
  • the encapsulation efficiency of the payload may be at least 70%.
  • 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%).
  • the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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%).
  • the encapsulation efficiency of the payload may be at least 70%.
  • the lipid-based carrier 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).
  • the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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).
  • the encapsulation efficiency of the payload may be at least 70%.
  • the lipid-based carrier (or lipid nanoformulation) comprises (i) a TREM (e.g., a TREM described herein); (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 Attorney Docket No.: F2099-7037WO 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.
  • a TREM e.g., a TREM described herein
  • a cationic lipid comprising from 50 mol% to 65 mol% of the
  • the lipid-based carrier (or lipid nanoformulation) comprises (i) a TREM (e.g., a TREM described herein); (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 carrier; and (d) a conjugated lipid comprising from 0.5 mol % to 2 mol % of the total lipid present in the lipid- based carrier.
  • a TREM e.g., a TREM described herein
  • a cationic lipid comprising from 50 mol % to 85 mol % of the total lipid present in the lipid-based carrier
  • a non-cationic lipid comprising from 13 mol % to 49.5 mol % of the total lipid present in the lipid-based carrier
  • 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.
  • the lipid-based carrier or lipid nanoformulation
  • 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.
  • the non-ionizable lipid components in the lipid-based carrier 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.
  • the ratio of total lipid components to the payload e.g., an encapsulated therapeutic agent such as a TREM (e.g., a TREM described herein) can be varied as desired.
  • the total lipid components to the payload (mass or weight) ratio can be from about 10:1 to about 30:1.
  • the total lipid components to the payload 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 payload 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, Attorney Docket No.: F2099-7037WO 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or higher.
  • the lipid-based carrier (or lipid nanoformulation’s) overall lipid content can range from about 5 mg/ml to about 30 mg/mL.
  • Nitrogen:phosphate ratios (N:P ratio) is evaluated at values between 0.1 and 100.
  • the efficiency of encapsulation of a payload such as a protein and/or nucleic acid describes the amount of protein and/or nucleic acid 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 protein or nucleic acid 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 protein or nucleic acid (e.g., RNA) in a solution.
  • Fluorescence may be used to measure the amount of free protein and/or nucleic acid (e.g., RNA) in a solution.
  • 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%.
  • the encapsulation efficiency may be at least 70%.
  • the encapsulation efficiency may be at least 80%.
  • the encapsulation efficiency may be at least 90%.
  • the encapsulation efficiency may be at least 95%.
  • TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition described herein can be administered to a cell without a carrier, e.g., via naked delivery of the TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition.
  • naked delivery as used herein refers to delivery without a carrier.
  • delivery without a carrier e.g., naked delivery, comprises delivery with a moiety, e.g., a targeting peptide.
  • a TREM, a TREM core fragment or a TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein is delivered to a cell without a carrier, e.g., via naked delivery.
  • the delivery without a carrier e.g., naked delivery, comprises delivery with a moiety, e.g., a targeting peptide.
  • a TREM composition e.g., a pharmaceutical TREM composition described herein
  • a TREM composition (e.g., a pharmaceutical TREM composition) described herein is contacted with a cell or tissue, or administered to a subject in need thereof, in an amount and for a time sufficient to modulate (increase or decrease) one or more of the following parameters: adaptor function (e.g., cognate or non-cognate adaptor function), e.g., the rate, efficiency, robustness, and/or specificity of initiation or elongation of a polypeptide chain; ribosome binding and/or occupancy; regulatory function (e.g., gene silencing or signaling); cell fate; mRNA stability; protein stability; protein transduction; protein compartmentalization.
  • adaptor function e.g., cognate or non-cognate adaptor function
  • regulatory function e.g., gene silencing or signaling
  • a parameter may be modulated, e.g., by at least 5% (e.g., at least 10%, 15%, 20%, 25%, 30%, 40%.50%.60%.70%, 80%, 90%, 100%, 150%, 200% or more) compared to a reference tissue, cell or subject (e.g., a healthy, wild-type or control cell, tissue or subject).
  • a reference tissue, cell or subject e.g., a healthy, wild-type or control cell, tissue or subject.
  • a tRNA effector molecule comprising a sequence of Formula (A): [L1]x-[ASt Domain1]-[L2]x-[DH Domain]-[L3]x-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]x-[ASt Domain2]-[L5]x (A), wherein: independently, [L1], [L5], and [VL Domain], are optional; x is 0 or 1; a nucleotide within the TREM sequence comprises a locked nucleic acid (LNA) moiety or 2’,5’-linked nucleotide.
  • LNA locked nucleic acid
  • the TREM of embodiment 1, wherein the LNA moiety comprises a heterocyclyl ring or cycloalkyl ring between the 2’-position and the 4’-position of the nucleotide sugar within the nucleotide. 3.
  • the TREM of embodiment 2, wherein the heterocyclyl ring or cycloalkyl ring is a five- membered ring, six-membered ring, seven-membered ring, or eight-membered ring. 5.
  • a naturally occurring nucleobase e.g., adenine, cytosine, guanine, uracil
  • B is a non-naturally occurring nucleobase.
  • B comprises a chemical modification (e.g.
  • the TREM of embodiment 8, wherein the chemical modification is a 2’OMe modification.
  • the TREM of embodiment 8, wherein the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification.
  • the TREM of embodiment 8, wherein the chemical modification is a 2’MOE modification.
  • the TREM of embodiment 8, wherein the chemical modification is a 2’-deoxy modification.
  • the TREM of embodiment 8, wherein the chemical modification is present in the internucleotide region (e.g., a backbone modification). 16.
  • the TREM of embodiment 8, wherein the chemical modification is a phosphorothioate modification. 17.
  • R 1 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • R 3 is hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • each of R 4a and R 4b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • the TREM of any one of embodiments 5-27, wherein one of L, M, Z, and X is N(R B )-. 29.
  • TREM any one of the preceding embodiments, wherein TREM comprises a single LNA moiety.
  • TREM comprises a plurality of LNA moieties.
  • the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 LNA moieties.
  • the LNA moiety is present in the [L1]-[ASt Domain1]-[L2].
  • 38. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. 39.
  • the TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L2]-[DH Domain]-[L3]. 40. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L3]-[ACH Domain]. 41. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [VL Domain]. Attorney Docket No.: F2099-7037WO 42. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [TH Domain]. 43. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L4]-[ASt Domain2]-[L5]. 44.
  • the TREM of any one of the preceding embodiments wherein the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, LNA, or phosphorothiorate modifications).
  • the TREM comprises the nucleotide sequence of any one of SEQ ID NOs: 626-793. 49.
  • each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene
  • the TREM of embodiment 49, wherein Y is selected from O or N(R B ). 51.
  • R 2 is hydrogen, C1-C6 alkyl, C1- C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. 55.
  • the TREM of any one of embodiments 49-61, wherein one of L and X is O.
  • each of R ia and R ib is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • each of Rii and Riii is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, or halo.
  • each of R ii and R iii is independently hydrogen.
  • Y is C(R’), O, or N(R B ); each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C1-
  • TREM of embodiment 71 wherein Y is selected from O or N(R B ).
  • Y is O.
  • 74 The TREM of any one of embodiments 71-73, wherein R 1 is hydrogen, C 1 -C 6 alkyl, C 1 - C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • 75 The TREM of any one of embodiments 71-74, wherein R 1 is hydrogen.
  • R 3 is hydrogen, C 1 -C 6 alkyl, C 1 - C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • X is selected from C1-C6 alkylene, -O-, -N(R B )-, -S-, and -Se-. 88.
  • each of R ia and R ib is independently hydrogen. 91.
  • each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or halo.
  • each of R ii and R iii is independently hydrogen.
  • TREM any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-c): tautomer, or hydrate thereof, wherein: Y is C(R’), O, or N(R B ); each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 hetero
  • R 1 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or halo.
  • each of R ii and R iii is independently hydrogen.
  • each of R 1 , R 2 , and R 3 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of L and Z is independently C 1 -C 6 alkylene, C 2 -C
  • R 1 is hydrogen, C1-C6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • R 3 is hydrogen, C1-C6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • the TREM of any one of embodiments 115-126, wherein one of L, M, Z, and X is N(R B )-. 128.
  • each of R ii and R iii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or halo. 136.
  • R 1 is hydrogen.
  • R 2 is hydrogen, C1-C6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • R 2 is hydrogen, C1-C6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • R 3 is hydrogen, C 1 -C 6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • R 3 is hydrogen, C 1 -C 6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • n is selected from 1, 2, or 3. 155.
  • TREM of any one of the preceding embodiments wherein n is 1. 156.
  • the TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-e-i) or (II-e-ii): e-ii) Attorney Docket No.: F2099-7037WO 157.
  • the TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety having a structure of Formula (II-f) is selected from: i), Attorney Docket No.: F2099-7037WO 160.
  • each of R 1 , R 2 , and R 3 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 4a and R 4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 1 -C 6 heteroalkyl, C 1
  • each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ;
  • X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(R B )-, -S(R D )x-, -Se(R E )
  • each of R 1 , R 2 , and R 3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 4a and R 4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl;
  • the TREM of one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety having a structure of Formula (II-l) is selected from: 166.
  • the TREM of one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-m-i): tautomer, or hydrate thereof. 167.
  • TREM any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-r): Attorney Docket No.: F2099-7037WO or hydrate thereof, wherein: a Y is C(R’), O, or N(R B ); each of R 1 , R 2 , and R 3 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; X is C 1 -C 6 alkylene, C 2 -C 6 alkenylene, C
  • TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety having a structure of Formula (II-r), is selected from Formula (II-s) or Formula (II-t): t), 169.
  • each of R 1 , R 2 , and R 4 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 3a and R 3b is independently hydrogen, C
  • a chemical modification e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification.
  • the TREM of embodiment 172, wherein the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. 174.
  • the TREM of embodiment 174, wherein the chemical modification is a 2’OMe modification.
  • the TREM of embodiment 174, wherein the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification.
  • the TREM of embodiment 174, wherein the chemical modification is a 2’MOE modification. 177.
  • the TREM of embodiment 174, wherein the chemical modification is a 2’-deoxy modification.
  • the TREM of embodiment 173, wherein the chemical modification is present in the internucleotide region (e.g., a backbone modification).
  • the TREM of embodiment 173, wherein the chemical modification is a phosphorothioate modification.
  • 180. The TREM of any one of embodiments 169-179, wherein the compound of Formula (III) does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification).
  • Y is O. Attorney Docket No.: F2099-7037WO 182.
  • R 2 is hydrogen, C1-C6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • R 2 is hydrogen, C1-C6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • R 4 is hydrogen, C 1 -C 6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • R 4 is hydrogen.
  • each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • 189 The TREM of any one of embodiments 169-188, wherein each of R 3a and R 3b is independently hydrogen. 190.
  • TREM any one of the preceding embodiments, wherein TREM comprises a single 2’,5’-linked nucleotide moiety. 191.
  • the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 252’,5’-linked nucleotides.
  • the TREM of any one of the preceding embodiments wherein the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications).
  • the TREM comprises the nucleotide sequence of 205.
  • each of R 1 , R 2 , and R 4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; R 3a is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6
  • the TREM of embodiment 205 wherein Y is O. 207.
  • the TREM of any one of embodiments 205-212, wherein R 3a is hydrogen, C 1 -C 6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo.
  • each of R 5a and R 5b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. 216.
  • the TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-a-i) or Formula (III-a-ii): a-ii) 218.
  • the TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-b):
  • each of R ia and R ib is independently hydrogen.
  • each of R ii and R iii is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl.
  • each of R ii and R iii is independently hydrogen.
  • each of R ia and R ib is independently hydrogen. 248.
  • each of R 1 , R 2 , and R 4 is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, C 2 -C 6 alkenyl,
  • each of R 3a and R 3b is independently hydrogen, C 1 -C 6 alkyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, or halo.
  • each of R ia and R ib is independently hydrogen. 266.
  • the TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-e):
  • each of R 1 , R 2 , and R 4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 haloalkyl, halo, cyano, -OR A , or -N(R B )(R C ), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R 10 ; each of R 3a and R 3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C 1 -C 6 heteroalkyl, C 1 -C 6 halo
  • a pharmaceutical composition comprising a TREM of any one of the preceding embodiments. 285.
  • a lipid nanoparticle formulation comprising a TREM of any one of embodiments 1-283. 287.
  • a lipid nanoparticle formulation comprising a pharmaceutical composition of embodiment 284. 288.
  • a method of treating a subject having a disease or disorder associated with a PTC comprising administering to the subject a TREM, TREM core fragment, or TREM fragment described herein (e.g., a TREM of any one of embodiments 1-283), thereby treating the subject having the disease or disorder.
  • the method of embodiment 288, wherein the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease.
  • TREM molecules e.g., modified TREMs
  • LNA locked nucleic acid
  • 2’,5’-linked nucleotide described herein may be chemically synthesized and purified by HPLC according to standard solid phase synthesis methods using phosphoramidite chemistry (see, e.g., Scaringe S. et al. (2004) Curr Protoc Nucleic Acid Chem, 2.10.1-2.10.16; Usman N. et al.
  • TREMs may also contain additional non-naturally occurring modifications, such as a 2’-methoxy (2’OMe), 2’fluoro (2’F), 2’-methoxyethyl (2’-MOE), or phosphorothioate (PS) modification.
  • arginine non- cognate TREM molecule named TREM-Arg-TGA contains the sequence of ARG-UCU-TREM body but with the anticodon sequence corresponding to UCA instead of UCU (i.e., SEQ ID NO: 622).
  • the following exemplary phosphoramidites were used: 5’-dimethoxytrityl-5-methyl-(2’-O, 4’-C methylene)-uridine,3’-[(2-cyanoethyl)-(N,N- diisopropyl)]-phosphoramidite, 5’-dimethoxytrityl-N-benzoyl-(2’-O, 4’-C methylene)- adenosine,3’-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 5’-dimethoxytrityl-N- dimethylformamidine-(2’-O, 4’-C methylene)-guanosine, 3’-[(2-cyanoethyl)-(N,N-diisopropyl)]- phosphoramidite, and 5’-dimethoxytrityl-N-
  • the following exemplary phosphoramidites were used: 5'-O-DMT-3'-TBDMS-N6-Bz-Adenosine 2'-CE phosphoramidite, 5'-O-DMT-3'-TBDMS-N4-Acetyl-Cytidine 2'-CE phosphoramidite, 5'-O-DMT-3'-O-TBDMS- N2-iBu-Guanosine 2'-CE phosphoramidite, and 5'-O-DMT-3'-TBDMS-Uridine 2'-CE phosphoramidite.
  • the phosphorothioate was introduced by oxidizing the phosphite triester using a sulfur transfer reagent, such as tetraethylthiuram disulfide (TETD), bis(O,O-diisopropoxy phosphinothioyl) disulfide (Stec’s reagent), 3H-1,2-benzodithiol-3-one-1,1,-dioxide (Beaucage reagent), phenylacetyl disulfide (PADS), 3-ethoxy-1,2,4-dithiazoline-5-one (EDITH), 1,2-dithiazole-5- thione (xanthane hydride or ADTT), 3-((dimethylamino-methylidene)amino)-3H-1,2,4- dithiazole-3-thione (DDTT), dimethylthiuram disulfide (DTD),
  • a sulfur transfer reagent such as tetraethylthiuram
  • FIG.1 describes a series of singly and multiply modified TREMs synthesized according to this procedure.
  • the sequences of each of these TREMs are provided in the table, wherein r: ribonucleotide; LNA: locked nucleic acid; [25r] or 2_5_r: 2’,5’-linked nucleotide; m: 2’-OMe; *: PS linkage; f: 2’-fluoro; moe: 2’-moe; d: deoxyribonucleotide; [m5C] or 5MeC: 5- methylcytosine.
  • mA represents 2’-O-methyl adenosine
  • moe5MeC represents 2’-MOE nucleotide with 5-methylcytosine nucleobase
  • dA represents an adenosine deoxyribonucleotide.
  • Example 2 HPLC and MS analysis of modified TREMs Attorney Docket No.: F2099-7037WO Chemically modified TREM molecules may be analyzed by HPLC, for example, to evaluate the purity and homogeneity of the compositions.
  • a Waters Aquity UPLC system using a Waters BEH C18 column (2.1 mm x 50 mm x 1.7 ⁇ m) may be used for this analysis.
  • Samples may be prepared by dissolving 0.5 nmol of the TREM in 75 ⁇ L of water and injecting 2 ⁇ L of the solution.
  • the buffers used may be 50 mM dimethylhexylammonium acetate with 10% CH3CN (acetonitrile) as buffer A and 50 mM dimethylhexylammonium acetate with 75% CH 3 CN as buffer B (gradient 25-75% buffer B over 5 mins), with a flow rate of 0.5 mL/min at 60 °C.
  • ESI-LCMS data for the chemically modified TREMs may be acquired on a Thermo Ultimate 3000-LTQ-XL mass spectrometer.
  • FIG.1 describe a series of singly and multiply modified TREMs synthesized according to the protocol outlined in Example 1. The calculated and detected molecular weights for each sequence were determined as outlined herein.
  • Example 3 Analysis of modified TREMs via anion-exchange HPLC This example describes the quality control of a synthesized TREM via anion-exchange HPLC. Using the Dionex DNA-Pac–PA-100 column, a gradient is employed using HPLC buffer A and HPLC buffer B.0.5 ODUs of a sample that has been dissolved in H2O or Tris buffer, pH 7.5 is injected onto the gradient. The gradient employed is based on oligonucleotide length and can be applied according to Table 13.
  • Table 14 The parameters provided in Table 14 can be used to program a linear gradient on the HPLC analyzer.
  • Table 13 Oligonucleotide length and gradient percentages Length Gradient
  • Table 14 Parameters for a linear gradient on HPLC analyzer Attorney Docket No.: F2099-7037WO Time Flow % Buffer % Buffer (min) (mL/min)
  • a B Analysis of TREMs via PAGE Purification and Analysis.
  • Gel purification and analysis of 2’-ACE protected RNA follows standard protocols for denaturing PAGE (Ellington and Pollard (1998) In Current Protocols in Molecular Biology, Chanda, V).
  • the 2’-ACE protected oligo is resuspended in 200 mL of gel loading buffer.
  • InvitrogenTM NuPAGETM 4-12% Bis-Tris Gels or similar gel is prepared in gel apparatus. Samples are loaded and gel ran at 50-120 W, maintaining the apparatus at 40°C. When complete, the gel is exposed to ultraviolet (UV) light at 254 nm to visualize the purity of the RNA using UV shadowing. If necessary, the desired gel band is excised with a clean razor blade. The gel slice is crushed and 0.3M NaOAc elution buffer is added to the gel particles, and soaked overnight. The mixture is decanted and filtered through a Sephadex column such as Nap-10 or Nap-25.
  • UV ultraviolet
  • Example 5 Deprotection of synthesized TREM This example describes the deprotection of a TREM made according to an in vitro synthesis method. The 2’-protecting groups are removed using 100 mM acetic acid, pH 3.8. The formic acid and ethylene glycol byproducts are removed by incubating at 60°C for 30 min Attorney Docket No.: F2099-7037WO followed by lyophilization or SpeedVac-ing to dryness. After this final deprotection step, the oligonucleotides are ready for use.
  • Example 6 Deprotection of synthesized TREM
  • Characterization of chemically modified TREMs for readthrough of a premature termination codon (PTC) in a reporter protein This example describes an assay to test the ability of a non-cognate chemically modified TREM to readthrough a PTC in a cell line expressing a reporter protein having a PTC.
  • This protocol describes analysis of chemically modified arginine and glutamate non-cognate TREM (i.e., Arg-TGA, Glu-TAG), though a non-cognate TREM specifying any one of the other amino acids can also be used.
  • a cell line engineered to stably express the NanoLuc reporter construct containing a premature termination codon (PTC) may be generated using the FlpIn and/or JumpIN cell system (ThermoFisher Scientific, USA) according to the manufacturer’s instructions.
  • the NanoLuc reporter can also be modified with a destabilizing PEST domain at its C-terminus to reduce the protein half-life.
  • Delivery of the chemically modified TREMs into the NanoLuc reporter cells is carried out via a reverse transfection reaction using lipofectamine RNAiMAX (ThermoFisher Scientific, USA) according to manufacturer instructions. Briefly, 0.15uL of a 5uM solution of chemically modified TREM sample are diluted in a 7.35uL RNAiMAX/OptiMEM mixture.
  • the 7.5uL TREM/transfection mixture is added to a 384-well plate and kept still for 20-30min before adding the cells stably expressing the NanoLuc reporter construct containing a PTC.
  • the NanoLuc reporter cells are harvested and diluted to 4 ⁇ 10 5 cells/mL in complete growth medium, and 30uL of the diluted cell suspension is added and mixed to the plate containing the TREM.
  • a NanoGlo bioluminescent assay may be performed according to manufacturer instruction.
  • NanoGlo reagent is prepared by mixing the buffer with substrate in a 50:1 ratio.15uL Attorney Docket No.: F2099-7037WO of mixed NanoGlo reagent is added to each well of the 384-well plate and mixed on the shaker at 600rpm for 10min. Then the plate is centrifuged at 100g for 1min, followed by a 5min incubation step at room temperature before measuring sample bioluminescence.
  • a host cell expressing the NanoLuc reporter construct with a PTC is used and transfected with a previously-identified unmodified, active TREM.
  • a negative control a host cell expressing the NanoLuc reporter construct with a PTC is used, but no TREM is transfected.
  • the efficacy of the chemically modified TREMs is measured as a ratio of the NanoLuc luminescence in the experimental sample to the NanoLuc luminescence of the positive control or as a ratio of the NanoLuc luminescence in the experimental sample to the NanoLuc luminescence of the negative control. It is expected that if the sample TREM is functional, it may be able to read-through the stop mutation in the NanoLuc reporter and produce a luminescent reading higher than the luminescent reading measured in the negative control. If the sample TREM is not functional, the stop mutation is not rescued, and luminescence less or equal to the negative control is detected.
  • the impacts of chemical modifications were evaluated in singly and multiply modified TREM sequences and are summarized in FIG.1.
  • the TREMs are annotated as follows: r: ribonucleotide; m: 2’-OMe; *: PS linkage; f: 2’-fluoro; moe: 2’-moe; d: deoxyribonucleotide; 5MeC: 5-methylcytosine.
  • mA represents 2’-O-methyl adenosine
  • moe5MeC represents 2’-MOE nucleotide with 5-methylcytosine nucleobase
  • dA represents an adenosine deoxyribonucleotide.
  • FIG.1 also summarizes the results of the activity screen in column “A” for measurements made using NanoLuc reporter cells at 48 hours post-transfection, which reported as log2 fold changes compared with the appropriate unmodified TREM, wherein “1” indicates less than a 1 log2 fold change; “2” indicates greater than or equal to 1 and less than 3.32 log2 fold change; and “3” indicates greater than or equal to 3.32 log2 fold change.
  • Example 7 Characterization of chemically modified TREMs for readthrough of a premature termination codon (PTC) in a disease reporter cell line
  • PTC premature termination codon
  • Host cell modification Attorney Docket No.: F2099-7037WO
  • PTC premature termination codon
  • Jump-In GripTite HEK293 (Thermo Scientific A14150) cells were co- transfected with an expression vector containing the disease reporter, such as pJTI-R4-DEST- CMV-FIX-R298X-HaloTag-HiBiT-pA for FIX R298X to make the Factor IX disease reporter expressing cell line, and a pJTI-R4-Int PhiC31 integrase expression vector using Lipofectamine2000 according to manufacturer’s instructions. After 24 hours, the media was replaced with fresh media.
  • the disease reporter such as pJTI-R4-DEST- CMV-FIX-R298X-HaloTag-HiBiT-pA for FIX R298X to make the Factor IX disease reporter expressing cell line
  • a pJTI-R4-Int PhiC31 integrase expression vector using Lipofectamine2000 according to manufacturer’s instructions. After 24 hours, the media was
  • TREMs were synthesized and characterized as described herein, then transfected into cells. Forty-eight hours after TREM delivery into cells, conditioned media was collected, fresh media was added to the cells and allowed to equilibrate to room temperature. To measure the efficacy of arginine TREMs in PTC readthrough, full-length HiBiT-tagged disease reporter protein was assayed in both cells, and 48-hour conditioned media.
  • Nano-Glo® HiBiT Lytic Reagent was added to both cells containing fresh media and 48-hour conditioned media at a 1:1 v/v ratio, mixed on an orbital shaker at 500rpm for 10 minutes, and incubated at room temperature for 10 minutes.
  • the HiBiT-tagged disease reporter activity is measured by reading the luminescence in a plate reader. The data is binned and quantified according to the scheme provided in Example 1.

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Abstract

The invention relates generally to tRNA-based effector molecules comprising a locked nucleotide acid (LNA) moiety, as well as compositions and methods relating thereto.

Description

Attorney Docket No.: F2099-7037WO MODIFIED TREMS, COMPOSITIONS, AND RELATED METHODS THEREOF CLAIM OF PRIORITY This application claims priority to U.S. Application No.63/458,793, filed April 12, 2023, and U.S. Application No.63/458,885, filed April 12, 2023. The disclosure of each of the foregoing applications is incorporated herein by reference in its entirety. BACKGROUND Transfer RNAs (tRNAs) are complex, naturally occurring RNA molecules that possess a number of functions including initiation and elongation of proteins. SUMMARY The present disclosure features modified tRNA-based effector molecules (TREMs, e.g., a TREM or TREM fragment), as well as related compositions and uses thereof. As provided herein, TREMs are complex molecules which can mediate a variety of cellular processes. The TREMs disclosed herein comprise a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, e.g., on a component nucleotide (e.g., a nucleotide sugar). In one aspect, provided herein is a TREM comprising a sequence of Formula (A): [L1]x-[ASt Domain1]-[L2]x- [DH Domain]-[L3]x-[ACH Domain]-[VL Domain]-[TH Domain]-[L4]x-[ASt Domain2]-[L5]x, wherein independently, [L1] and [VL Domain], are optional; x, independently for each occurrence, is 0 or 1; and one or more of [L1], [ASt Domain1], [L2]-[DH Domain], [L3], [ACH Domain], [VL Domain], [TH Domain], [L4], [ASt Domain2], and [L5] comprises a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide. In an embodiment, the TREM comprises a sequence of Formula (A-1): [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]- [ACH Domain]-[VL Domain]-[TH Domain]-[L4]-[ASt Domain2], wherein independently, [L1] and [VL Domain], are optional; wherein one of [L1], [ASt Domain1], [L2]-[DH Domain], [L3], [ACH Domain], [VL Domain], [TH Domain], [L4], and [ASt Domain2] comprises a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide. In an embodiment, the TREM further comprises an additional non-naturally occurring modification. In some embodiments, the TREMs disclosed herein further comprise at least one additional modification Attorney Docket No.: F2099-7037WO (e.g., a non-naturally occurring modification), e.g., on a component nucleotide (e.g., a nucleobase or sugar) or within an internucleotide region, e.g., the TREM backbone. In an embodiment, the TREM: (a) has the ability to: (i) support protein synthesis, (ii) be charged by a synthetase, (iii) be bound by an elongation factor, (iv) introduce an amino acid into a peptide chain, (v) support elongation, or (vi) support initiation; (b) comprises at least X contiguous nucleotides without a non-naturally occurring modification, wherein X is greater than 3, 4, 5, 6, 7, 8, 9, or 10; (c) comprises at least 3, but less than all of the nucleotides of a type (e.g., A, T, C, G or U) comprise the same non-naturally occurring modification; (d) comprises at least X nucleotides of a type (e.g., A, T, C, G or U) that do not comprise a non-naturally occurring modification, wherein X= than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, or 80; (e) comprises no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, or 80 nucleotides of a type (e.g., A, T, C, G or U) that comprise a non-naturally occurring modification; and/or (f) comprises no more than than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, or 80 nucleotides of a type (e.g., A, T, C, G or U) that do not comprise a non- naturally occurring modification. In an embodiment, the TREM comprises feature (a)(i). In an embodiment, the TREM comprises feature (a)(ii). In an embodiment, the TREM comprises feature (a)(iii). In an embodiment, the TREM comprises feature (a)(iv). In an embodiment, the TREM comprises feature (a)(v). In an embodiment, the TREM comprises feature (a)(vi). In an embodiment, the TREM comprises feature (b). In an embodiment, the TREM comprises feature (c). In an embodiment, the TREM comprises feature (d). In an embodiment, the TREM comprises feature (e). In an embodiment, the TREM comprises feature (f). In an embodiment, the TREM comprises all of features (a)-(f) or a combination thereof. In an embodiment, the TREM Domain comprising the non-naturally occurring modification has a function, e.g., a domain function described herein. In an aspect, provided herein is a TREM core fragment comprising a sequence of Formula B: Attorney Docket No.: F2099-7037WO [L1] y-[ASt Domain1] x-[L2] y-[DH Domain]y-[L3] y-[ACH Domain]x-[VL Domain] y-[TH Domain] y-[L4] y-[ASt Domain2] x, wherein x=1 and y=0 or 1; and one of [ASt Domain1], [ACH Domain], and [ASt Domain2] comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the TREM has the ability to support protein synthesis. In an embodiment, the TREM has the ability to be able to be charged by a synthetase. In an embodiment, the TREM has the ability to be bound by an elongation factor. In an embodiment, the TREM has the ability to introduce an amino acid into a peptide chain. In an embodiment, the TREM has the ability to support elongation. In an embodiment, the TREM has the ability to support initiation. In an embodiment, the [ASt Domain 1] and/or [ASt Domain 2] comprising a non- naturally occurring modification has the ability to initiate or elongate a polypeptide chain. In an embodiment, the [ACH Domain] comprising a non-naturally occurring modification has the ability to mediate pairing with a codon. In an embodiment, y=1 for any one, two, three, four, five, six, all or a combination of [L1], [L2], [DH Domain], [L3], [VL Domain], [TH Domain], [L4]. In an embodiment, y=0 for any one, two, three, four, five, six, all or a combination of [L1], [L2], [DH Domain], [L3], [VL Domain], [TH Domain], [L4]. In an embodiment, y=1 for linker [L1], and L1 comprises a nucleotide having a non- naturally occurring modification. In an embodiment, y=1 for linker [L2], and L2 comprises a nucleotide having a non- naturally occurring modification. In an embodiment, y=1 for [DH Domain (DHD)], and DHD comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the DHD comprising the non- naturally occurring modification has the ability to mediate recognition of aminoacyl-tRNA synthetase. In an embodiment, y=1 for linker [L3], and L3 comprises a nucleotide having a non- naturally occurring modification. In an embodiment, y=1 for [VL Domain (VLD)], and VLD comprises a nucleotide having a non-naturally occurring modification. Attorney Docket No.: F2099-7037WO In an embodiment, y=1 for [TH Domain (THD)], and THD comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the THD comprising the non- naturally occurring modification has the ability to mediate recognition of the ribosome. In an embodiment, y=1 for linker [L4], and L4 comprises a nucleotide having a non- naturally occurring modification. In another aspect, the disclosure provides a TREM fragment comprising a portion of a TREM, wherein the TREM comprises a sequence of Formula (A-1): [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2], and wherein the TREM fragment comprises a non-naturally occurring modification. In an embodiment, the TREM fragment comprises one, two, three or all or any combination of the following: (a) a TREM half (e.g., from a cleavage in the ACH Domain, e.g., in the anticodon sequence, e.g., a 5’half or a 3’ half); (b) a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DH Domain or the ACH Domain); (c) a 3’ fragment (e.g., a fragment comprising the 3’ end, e.g., from a cleavage in the TH Domain); or (d) an internal fragment (e.g., from a cleavage in any one of the ACH Domain, DH Domain or TH Domain). In an embodiment, the TREM fragment comprise (a) a TREM half which comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the TREM fragment comprise (b) a 5’ fragment which comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the TREM fragment comprise (c) a 3’ fragment which comprises a nucleotide having a non-naturally occurring modification. In an embodiment, the TREM fragment comprise (d) an internal fragment which comprises a nucleotide having a non-naturally occurring modification. In an embodiment of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the TREM Domain comprises a plurality of nucleotides each having a non- naturally occurring modification. In an embodiment, the non-naturally occurring modification comprises a nucleobase modification, a sugar (e.g., ribose) modification, or a backbone modification. In an embodiment, tbe non-naturally occurring modification is a sugar (e.g., ribose) modification. In an embodiment, tbe non-naturally occurring modification is 2’-ribose Attorney Docket No.: F2099-7037WO modification, e.g., a 2’-OMe, 2’-halo (e.g., 2’-F), 2’-MOE, or 2’-deoxy modification. In an embodiment, tbe non-naturally occurring modification is a backbone modification, e.g., a phosphorothioate modification. In an embodiment of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the TREM sequence comprises a CCA sequence on a terminus, e.g., the 3’ terminus. In an embodiment, the TREM sequence does not comprise a CCA sequence on a terminus, e.g., the 3’ terminus. In an embodiment of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the non-naturally occurring modification is a base modification chosen from a modification listed in Table 5. In an embodiment of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the TREM, TREM core fragment, or TREM fragment is encoded by a sequence provided in Table 1, e.g., any one of SEQ ID NOs 1-451. In an embodiment of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the TREM, TREM core fragment, or TREM fragment is encoded by a consensus sequence chosen from any one of SEQ ID NOs: 562-621. In an embodiment of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the TREM, TREM core fragment, or TREM fragment is encoded by a sequence provided in FIG.1, e.g., any one of SEQ ID NOs: 625-1416, e.g., SEQ ID NOs: 625-793, 794- 991, or 992-1416. In an embodiment, the TREM, TREM core fragment, or TREM fragment comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in FIG.1, e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416. In an embodiment, the TREM, TREM core fragment, or TREM fragment comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from a TREM provided in FIG.1, e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416. In an embodiment, the TREM, TREM core fragment, or TREM fragment comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non- naturally occurring modifications compared with a TREM, TREM core fragment, or TREM fragment provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications), e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416. In an Attorney Docket No.: F2099-7037WO embodiment, the TREM, TREM core fragment, or TREM fragment comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional LNA moieties or 2’,5’-linked nucleotides compared with a TREM, TREM core fragment, or TREM fragment provided in FIG.1, e.g., any one of SEQ ID Nos: 625-1416, e.g., any one of SEQ ID NOs: 625-793, 794-991, or 992-1416. In an embodiment of any of the TREMs, TREM core fragments, or TREM fragments disclosed herein, the TREM, TREM core fragment, or TREM fragment is a TREM provided in FIG.1, e.g., any one of TREM NOs: 1-791, e.g., any one of TREM NOs: 1-169, 170-367, or 368-791. In an embodiment, the TREM, TREM core fragment, or TREM fragment comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications), e.g., any one of TREM NOs: 1-791, e.g., any one of TREM Nos: 1-169, 170-367, or 368-791. In another aspect, the disclosure provides a pharmaceutical composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein. In another aspect, a TREM or a related composition thereof can be used, inter alia, to modulate a functional parameter (e.g., an expression parameter and/or a signaling parameter) of an RNA corresponding to, or a polypeptide encoded by, a nucleic acid sequence comprising an endogenous open reading frame (ORF) having a premature termination codon (PTC). In another aspect, provided herein is a method of modulating a functional parameter of an mRNA corresponding to, or polypeptide encoded by, an endogenous open reading frame (ORF) in a subject, which ORF comprises a premature termination codon (PTC), contacting the subject with a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein (e.g., a TREM, TREM core fragment, or a TREM fragment comprising an LNA moiety or a 2’,5’-linked nucleotide) in an amount and/or for a time sufficient to modulate the functional parameter of the mRNA or polypeptide, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the codon having the first sequence, thereby modulating the functional parameter in the subject. In an embodiment, the functional parameter comprises a signaling parameter and/or an expression parameter, e.g., as described herein. In another aspect, disclosed herein is a method of modulating expression of a protein in a cell, wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), which ORF comprises a premature termination codon (PTC), comprising Attorney Docket No.: F2099-7037WO contacting the cell with a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein (e.g., a TREM, TREM core fragment, or a TREM fragment comprising an LNA moiety or a 2’,5’-linked nucleotide) in an amount and/or for a time sufficient to modulate expression of the encoded protein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the PTC, thereby modulating expression of the protein in the cell. In an embodiment, the PTC comprises UAA, UGA or UAG. In another aspect, provided herein is a method of increasing expression of a protein in a subject wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF), which ORF comprises a premature termination codon (PTC), comprising contacting the subject, in an amount and/or for a time sufficient to increase expression of the protein, with a TREM composition that (i) has an anticodon that pairs with the PTC, (ii) recognizes an aminoacyl-tRNA synthetase specific for Trp, Tyr, Cys, Glu, Lys, Gln, Ser, Leu, Arg, or Gly, (iii) comprises a sequence of Formula A, or (iv) comprises a non-naturally occurring modification. In an embodiment, the PTC comprises UAA, UGA or UAG. In an embodiment, the TREM composition comprises (i). In an embodiment, the TREM composition comprises (ii). In an embodiment, the TREM composition comprises (iii). In an embodiment, the TREM composition comprises (iv). In an embodiment, the TREM composition comprises two of (i)- (iv). In an embodiment, the TREM composition comprises three of (i)-(iv). In an embodiment, the TREM composition comprises each of (i)-(iv). In another aspect, the disclosure provides a method of treating a subject having an endogenous open reading frame (ORF) which comprises a premature termination codon (PTC), comprising providing a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM comprises an anticodon that pairs with the PTC in the ORF; contacting the subject with the composition comprising a TREM, TREM core fragment or TREM fragment in an amount and/or for a time sufficient to treat the subject, thereby treating the subject. In an embodiment, the PTC comprises UAA, UGA or UAG. In another aspect, the disclosure provides a method of treating a subject having an disease or disorder associated with a premature termination codon (PTC), comprising providing a TREM composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein (e.g., a TREM, TREM core fragment, or a TREM fragment comprising an LNA moiety or a 2’,5’-linked nucleotide); contacting the subject with the composition comprising a TREM, Attorney Docket No.: F2099-7037WO TREM core fragment or TREM fragment in an amount and/or for a time sufficient to treat the subject, thereby treating the subject. In an embodiment, the PTC comprises UAA, UGA or UAG. In an embodiment, the disease or disorder associated with a PTC is a disease or disorcer described herein, e.g., a cancer or a monogenic disease. In an embodiment of any of the methods disclosed herein, the codon having the first sequence comprises a mutation (e.g., a point mutation, e.g., a nonsense mutation), resulting in a premature termination codon (PTC) chosen from UAA, UGA or UAG. In an embodiment, the codon having the first sequence or the PTC comprises a UAA mutation. In an embodiment, the codon having the first sequence or the PTC comprises a UGA mutation. In an embodiment, the codon having the first sequence or the PTC comprises a UAG mutation In another aspect, the disclosure provides a method of making a TREM, a TREM core fragment, or a TREM fragment disclosed herein, comprising linking a first nucleotide to a second nucleotide to form the TREM. In an embodiment, the TREM, TREM core fragment or TREM fragment is non-naturally occurring (e.g., synthetic). In an embodiment, the TREM, TREM core fragment or TREM fragment is made by cell- free solid phase synthesis. In another aspect, the disclosure provides a method of modulating a tRNA pool in a cell comprising: providing a TREM, a TREM core fragment, or a TREM fragment disclosed herein, and contacting the cell with the TREM, TREM core fragment or TREM fragment, thereby modulating the tRNA pool in the cell. In an aspect, the disclosure provides a method of contacting a cell, tissue, or subject with a TREM, a TREM core fragment, or a TREM fragment disclosed herein, comprising: contacting the cell, tissue or subject with the TREM, TREM core fragment or TREM fragment, thereby contacting the cell, tissue, or subject with the TREM, TREM core fragment or TREM fragment. In another aspect, the disclosure provides a method of delivering a TREM, TREM core fragment or TREM fragment to a cell, tissue, or subject, comprising: providing a cell, tissue, or subject, and contacting the cell, tissue, or subject, a TREM, a TREM core fragment, or a TREM fragment disclosed herein. Attorney Docket No.: F2099-7037WO In an aspect, the disclosure provides a method of modulating a tRNA pool in a cell comprising an endogenous open reading frame (ORF), which ORF comprises a codon having a first sequence, comprising: optionally, acquiring knowledge of the abundance of one or both of (i) and (ii), e.g., acquiring knowledge of the relative amounts of: (i) and (ii) in the cell, wherein (i) is a tRNA moiety having an anticodon that pairs with the codon of the ORF having a first sequence (the first tRNA moiety) and (ii) is an isoacceptor tRNA moiety having an anticodon that pairs with a codon other than the codon having the first sequence (the second tRNA moiety) in the cell; contacting the cell with a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with: the codon having the first sequence; or the codon other than the codon having the first sequence, in an amount and/or for a time sufficient to modulate the relative amounts of the first tRNA moiety and the second tRNA moiety in the cell, thereby modulating the tRNA pool in the cell. In another aspect, the disclosure provides a method of modulating a tRNA pool in a subject having an ORF, which ORF comprises a codon having a first sequence, comprising: optionally, acquiring knowledge of the abundance of one or both of (i) and (ii), e.g., acquiring knowledge of the relative amounts of: (i) and (ii) in the subject, wherein (i) is a tRNA moiety having an anticodon that pairs with the codon of the ORF having a first sequence (the first tRNA moiety) and (ii) is an isoacceptor tRNA moiety having an anticodon that pairs with a codon other than the codon having the first sequence (the second tRNA moiety) in the subject; contacting the subject with a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with: the codon having the first sequence; or the codon other than the codon having the first sequence, in an amount and/or for a time sufficient to modulate the relative amounts of the first tRNA moiety and the second tRNA moiety in the subject, thereby modulating the tRNA pool in the subject. In an aspect, the disclosure provides a method of modulating a tRNA pool in a subject having an endogenous ORF comprising a codon comprising a synonymous mutation (a synonymous mutation codon or SMC), comprising: Attorney Docket No.: F2099-7037WO providing a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment comprises an isoacceptor tRNA moiety comprising an anticodon sequence that pairs with the SMC (the TREM); contacting the subject with the composition in an amount and/or for a time sufficient to modulate the tRNA pool in the subject, thereby modulating the tRNA pool in the subject. In another aspect, the disclosure provides a method of modulating a tRNA pool in a cell comprising an endogenous ORF comprising a codon comprising a SMC, comprising: providing a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, wherein the TREM, TREM core fragment or TREM fragment comprises an isoacceptor tRNA moiety comprising an anticodon sequence that pairs with the SMC (the TREM); contacting the cell with the composition comprising a TREM in an amount and/or for a time sufficient to modulate the tRNA pool in the cell, thereby modulating the tRNA pool in the cell. In an aspect, the disclosure provides a method of modulating expression of a protein in a cell, wherein the protein is encoded by a nucleic acid comprising an ORF, which ORF comprises a codon having a mutation, comprising: contacting the cell with a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein in an amount and/or for a time sufficient to modulate expression of the encoded protein, wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the codon having the mutation, thereby modulating expression of the protein in the cell. In another aspect, the disclosure provides a method of modulating expression of a protein in a subject, wherein the protein is encoded by a nucleic acid comprising an endogenous ORF, which ORF comprises a codon having a mutation, comprising: contacting the subject with a composition comprising a TREM, a TREM core fragment, or a TREM fragment disclosed herein, in an amount and/or for a time sufficient to modulate expression of the encoded protein, Attorney Docket No.: F2099-7037WO wherein the TREM, TREM core fragment or TREM fragment has an anticodon that pairs with the codon having the mutation, thereby modulating expression of the protein in the subject. In an embodiment of any of the methods disclosed herein, the mutation in the ORF is a nonsense mutation, e.g., resulting in a premature stop codon chosen from UAA, UGA or UAG. In an embodiment, the stop codon is UAA. In an embodiment, the stop codon is UGA. In an embodiment, the stop codon is UAG. In an embodiment of any of the methods disclosed herein, the TREM comprises an anticodon that pairs with a stop codon. TREMs of the disclosure include TREMs, TREM core fragments and TREM fragments. TREMs, TREM core fragments or TREM fragments can be modified with non-naturally occurring modifications to, e.g., increase the level and/or activity (e.g., stability) of the TREM. Pharmaceutical TREM compositions, e.g., comprising TREMs having a non-naturally occurring modification, can be administered to cells, tissues or subjects to modulate these functions, e.g., in vitro or in vivo. Disclosed herein are TREMs, TREM core fragments or TREM fragments comprising non-naturally occurring modifications, TREM compositions, preparations, methods of making TREM compositions and preparations, and methods of using the same. In an embodiment, the TREM, TREM core fragment, and TREM fragments comprise a non-naturally occurring modification that improves stability or enhances activity of the TREM, TREM core fragment, or TREM fragment. Additional features of any of the aforesaid TREMs, TREM core fragments, TREM fragments, TREM compositions, preparations, methods of making TREM compositions and preparations, and methods of using TREM compositions and preparations include one or more of the features in the Enumerated Embodiments, Figures, Description, Examples, or Claims. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following Enumerated Embodiments, Drawings, Description, Examples, or Claims. BRIEF DESCRIPTION OF THE DRAWINGS Attorney Docket No.: F2099-7037WO FIG.1 is a listing of exemplary TREMs described herein, e.g., TREMs comprising a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide. DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS The present disclosure features tRNA-based effector molecules (TREMs) comprising a nucleotide comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, as well as compositions and related methods of use and making. As disclosed herein, TREMs are complex molecules which can mediate a variety of cellular processes. Pharmaceutical TREM compositions, e.g., TREMs comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, can be administered to a cell, a tissue, or to a subject to modulate these functions. Also disclosed herein are methods of modulating expression of a protein in a subject or cell, wherein the protein is encoded by a nucleic acid comprising an endogenous open reading frame (ORF) having a first sequence, e.g., a mutation, e.g., a premature termination codon (PTC), and methods of treating a subject having an endogenous open reading frame (ORF) which comprises a premature termination codon (PTC). Further disclosed herein are TREMs comprising a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide, and an additional non-naturally occurring modification, methods of making the same and compositions thereof. Definitions “Acquire” or “acquiring” as the terms are used herein, refer to obtaining possession of a value, e.g., a numerical value, by “directly acquiring” or “indirectly acquiring” the physical entity or value. “Directly acquiring” refers to performing a process (e.g., performing an analytical method) to obtain the value. “Indirectly acquiring” refers to receiving the value from another party or source (e.g., a third party laboratory that directly acquired the or value). A “disease or disorder associated with a PTC” as that term is used herein includes, but is not limited to, a disease or disorder in which cells express, or at one time expressed, a polypeptide encoded by an ORF comprising a PTC. In some embodiments, a disease associated with a PTC is chosen from: a proliferative disorder (e.g., a cancer), a genetic disorder, a metabolic disorder, an immune disorder, an inflammatory disorder or a neurological disorder. Exemplary diseases or disorders associated with a PTC are provided in any one of Tables 15, 16 Attorney Docket No.: F2099-7037WO and 17. In an embodiment, the disease associated with a PTC is a cancer. In an embodiment, the disease associated with a PTC is a monogenic disease. An “isoacceptor,” as that term is used herein, refers to a plurality of tRNA molecule or TREMs wherein each molecule of the plurality comprises a different naturally occurring anticodon sequence and each molecule of the plurality mediates the incorporation of the same amino acid and that amino acid is the amino acid that naturally corresponds to the anticodons of the plurality. A “modification,” as that term is used herein with reference to a nucleotide, refers to a modification of the chemical structure, e.g., a covalent modification, of the subject nucleotide. The modification can be naturally occurring or non-naturally occurring. In an embodiment, the modification is non-naturally occurring. In an embodiment, the modification is naturally occurring. In an embodiment, the modification is a synthetic modification. In an embodiment, the modification is a modification provided in Table 5. A “naturally occurring nucleotide,” as that term is used herein, refers to a nucleotide that does not comprise a non-naturally occurring modification. In an embodiment, it includes a naturally occurring modification. A “non-naturally occurring modification,” as that term is used herein with reference to a nucleotide, refers to a modification that: (a) a cell, e.g., a human cell, does not make on an endogenous tRNA; or (b) a cell, e.g., a human cell, can make on an endogenous tRNA but wherein such modification is in a location in which it does not occur on a native tRNA, e.g., the modification is in a domain, linker or arm, or on a nucleotide and/or at a position within a domain, linker or arm, which does not have such modification in nature. In either case, the modification is added synthetically, e.g., in a cell free reaction, e.g., in a solid state or liquid phase synthetic reaction. In an embodiment, the non-naturally occurring modification is a modification that is not present (in identity, location or position) if a sequence of the TREM is expressed in a mammalian cell, e.g., a HEK293 cell line. Exemplary non-naturally occurring modifications are found in Table 5. In an embodiment, the non-naturally occurring modification comprises a locked nucleic acid moiety or a 2’,5’-linked nucleotide. In an embodiment, the non- naturally occurring modification comprises a locked nucleic acid moiety. In an embodiment, the non-naturally occurring modification comprises a 2’,5’-linked nucleotide. Attorney Docket No.: F2099-7037WO A “non-naturally modified nucleotide,” as that term is used herein, refers a nucleotide comprising a non-naturally occurring modification on or of a sugar, nucleobase, or phosphate moiety. A “nucleotide,” as that term is used herein, refers to an entity comprising a sugar, typically a pentameric sugar; a nucleobase; and a phosphate linking group. In an embodiment, a nucleotide comprises a naturally occurring, e.g., naturally occurring in a human cell, nucleotide, e.g., an adenine, thymine, guanine, cytosine, or uracil nucleotide. A “premature termination codon” or “PTC” as those terms are used herein, refer to a stop codon that occurs in an open reading frame (ORF) of a DNA or mRNA. In an embodiment, a PTC occurs at a position upstream of a naturally occurring stop codon in an ORF. In an embodiment, a PTC that occurs upstream of a naturally occurring stop codon, e.g., in an ORF, results in modulation of a functional parameter of the corresponding mRNA or polypeptide encoded by the ORF. In an embodiment, a PTC can differ (or arise) from a pre-mutation sequence by a point mutation, e.g., a nonsense mutation. In an embodiment, a PTC can differ (or arise) from a pre-mutation sequence by a genetic change, e.g., abnormality, other than a point mutation, e.g., a frameshift, a deletion, an insertion, a rearrangement, an inversion, a translocation, a duplication, or a transversion. In an embodiment, a PTC results in the production of a truncated protein which lacks a native activity or which is associated with a mutant, disease, or other unwanted phenotype. In an embodiment, the ORF comprising the PTC is an ORF from a tumor suppressor gene. In an embodiment, the mutation giving rise to the PTC is a driver mutation, e.g., a mutation that provides a growth advantage to a tumor cell. A “functional parameter,” refers to an expression parameter and/or a signaling parameter. In an embodiment a functional parameter is an expression parameter. An expression parameter includes an expression parameter of a polypeptide or protein encoded by the endogenous ORF having a first sequence or PTC; or an expression parameter of an RNA, e.g., messenger RNA, encoded by the endogenous ORF having a first sequence or PTC. In an embodiment, an expression parameter can include: (a) protein translation; (b) expression level (e.g., of polypeptide or protein, or mRNA); (c) post-translational modification of polypeptide or protein; (d) folding (e.g., of polypeptide or protein, or mRNA), Attorney Docket No.: F2099-7037WO (e) structure (e.g., of polypeptide or protein, or mRNA), (f) transduction (e.g., of polypeptide or protein), (g) compartmentalization (e.g., of polypeptide or protein, or mRNA), (h) incorporation (e.g., of polypeptide or protein, or mRNA) into a supermolecular structure, e.g., incorporation into a membrane, proteasome, or ribosome, (i) incorporation into a multimeric polypeptide, e.g., a homo or heterodimer, and/or (j) stability. In an embodiment, a functional parameter is a signaling parameter. A signaling parameter can include: (1) modulation of a signaling pathway, e.g., a cellular signaling pathway which is downstream or upstream of the protein encoded by the endogenous ORF having a first sequence or PTC; (2) cell fate modulation; (3) ribosome occupancy modulation; (4) protein translation modulation; (5) mRNA stability modulation; (6) protein folding and structure modulation; (7) protein transduction or compartmentalization modulation; and/or (8) protein stability modulation. An “ORF having a PTC” as that phrase is used herein, refers to an open reading frame (ORF) which comprises a premature termination codon (PTC). In an embodiment, the ORF having the PTC is associated with a disease or disorder associated with a PTC, e.g., as described herein, e.g., a disease or disorder listed in any one of Tables 15, 16 and 17. In an embodiment, the ORF having the PTC is not associated with a disease or disorder associated with a PTC. A “stop codon” as that term is used herein, refers to a three nucleotide contiguous sequence within messenger RNA that specifies a termination of translation. For example, UAG, UAA, UGA (in RNA) and TAG, TAA or TGA (in DNA) are stop codons. The stop codons are also known as amber (UAG), ochre (UAA), and opal (UGA). A “tRNA-based effector molecule” or “TREM,” as that term is used herein, refers to an RNA molecule comprising a structure or property from (a)-(v) below, and which is a recombinant TREM, a synthetic TREM, or a TREM expressed from a heterologous cell. The Attorney Docket No.: F2099-7037WO TREMs described in the present disclosure comprise a nucleotide bearing a locked nucleic acid moiety or a 2’,5’-linked nucleotide. In addition, the TREM described herein are synthetic molecules and are made, e.g., in a cell free reaction, e.g., in a solid state or liquid phase synthetic reaction. TREMs are chemically distinct, e.g., in terms of primary sequence, type or location of modifications from the endogenous tRNA molecules made in cells, e.g., in mammalian cells, e.g., in human cells. A TREM can have a plurality (e.g., 2, 3, 4, 5, 6, 7, 8, 9) of the structures and functions of (a)-(v). In an embodiment, a TREM is non-native, as evaluated by structure or the way in which it was made. In an embodiment, a TREM comprises one or more of the following structures or properties: (a’) an optional linker region of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 1 region; (a) an amino acid attachment domain that binds an amino acid, e.g., an acceptor stem domain (AStD), wherein an AStD comprises sufficient RNA sequence to mediate, e.g., when present in an otherwise wildtype tRNA, acceptance of an amino acid, e.g., its cognate amino acid or a non-cognate amino acid, and transfer of the amino acid (AA) in the initiation or elongation of a polypeptide chain. Typically, the AStD comprises a 3’-end adenosine (CCA) for acceptor stem charging which is part of synthetase recognition. In an embodiment the AStD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring AStD, e.g., an AStD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of an AStD, e.g., an AStD encoded by a nucleic acid in Table 1, which fragment in embodiments has AStD activity and in other embodiments does not have AStD activity. (One of ordinary skill can determine the relevant corresponding sequence for any of the domains, stems, loops, or other sequence features mentioned herein from a sequence encoded by a nucleic acid in Table 1. E.g., one of ordinary skill can determine the sequence which corresponds to an AStD from a tRNA sequence encoded by a nucleic acid in Table 1.) In an embodiment the AStD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions; Attorney Docket No.: F2099-7037WO In an embodiment, the AStD comprises residues R1-R2-R3-R4 -R5-R6-R7 and residues R65- R66-R67-R68-R69-R70-R71 of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the AStD comprises residues R1-R2-R3-R4 -R5-R6-R7 and residues R65- R66-R67-R68-R69-R70-R71 of Formula II ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the AStD comprises residues R1-R2-R3-R4 -R5-R6-R7 and residues R65- R66-R67-R68-R69-R70-R71 of Formula III ZZZ, wherein ZZZ indicates any of the twenty amino acids; (a’-1) a linker comprising residues R8-R9 of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 2 region; (b) a dihydrouridine hairpin domain (DHD), wherein a DHD comprises sufficient RNA sequence to mediate, e.g., when present in an otherwise wildtype tRNA, recognition of aminoacyl-tRNA synthetase, e.g., acts as a recognition site for aminoacyl-tRNA synthetase for amino acid charging of the TREM. In embodiments, a DHD mediates the stabilization of the TREM’s tertiary structure. In an embodiment the DHD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring DHD, e.g., a DHD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a DHD, e.g., a DHD encoded by a nucleic acid in Table 1, which fragment in embodiments has DHD activity and in other embodiments does not have DHD activity. In an embodiment the DHD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions; In an embodiment, the DHD comprises residues R10-R11-R12-R13-R14 R15-R16-R17-R18- R19-R20-R21-R22-R23-R24-R25-R26-R27-R28 of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the DHD comprises residues R10-R11-R12-R13-R14 R15-R16-R17-R18- R19-R20-R21-R22-R23-R24-R25-R26-R27-R28 of Formula II ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the DHD comprises residues R10-R11-R12-R13-R14 R15-R16-R17-R18- R19-R20-R21-R22-R23-R24-R25-R26-R27-R28 of Formula III ZZZ, wherein ZZZ indicates any of the twenty amino acids; Attorney Docket No.: F2099-7037WO (b’-1) a linker comprising residue R29 of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 3 region; (c) an anticodon that binds a respective codon in an mRNA, e.g., an anticodon hairpin domain (ACHD), wherein an ACHD comprises sufficient sequence, e.g., an anticodon triplet, to mediate, e.g., when present in an otherwise wildtype tRNA, pairing (with or without wobble) with a codon; In an embodiment the ACHD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of an ACHD, e.g., an ACHD encoded by a nucleic acid in Table 1, which fragment in embodiments has ACHD activity and in other embodiments does not have ACHD activity. In an embodiment the ACHD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions; In an embodiment, the ACHD comprises residues -R30-R31-R32-R33-R34-R35-R36-R37-R38- R39-R40-R41-R42-R43-R44-R45-R46 of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the ACHD comprises residues -R30-R31-R32-R33-R34-R35-R36-R37-R38- R39-R40-R41-R42-R43-R44-R45-R46 of Formula II ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the ACHD comprises residues -R30-R31-R32-R33-R34-R35-R36-R37-R38- R39-R40-R41-R42-R43-R44-R45-R46 of Formula III ZZZ, wherein ZZZ indicates any of the twenty amino acids; (d) a variable loop domain (VLD), wherein a VLD comprises sufficient RNA sequence to mediate, e.g., when present in an otherwise wildtype tRNA, recognition of aminoacyl-tRNA synthetase, e.g., acts as a recognition site for aminoacyl-tRNA synthetase for amino acid charging of the TREM. In embodiments, a VLD mediates the stabilization of the TREM’s tertiary structure. In an embodiment, a VLD modulates, e.g., increases, the specificity of the TREM, e.g., for its cognate amino acid, e.g., the VLD modulates the TREM’s cognate adaptor function. In an embodiment the VLD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring VLD, e.g., a VLD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a VLD, e.g., a VLD encoded by a nucleic acid Attorney Docket No.: F2099-7037WO in Table 1, which fragment in embodiments has VLD activity and in other embodiments does not have VLD activity. In an embodiment the VLD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section. In an embodiment, the VLD comprises residue -[R47]x of a consensus sequence provided in the “Consensus Sequence” section, wherein x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1- 175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271); (e) a thymine hairpin domain (THD), wherein a THD comprises sufficient RNA sequence, to mediate, e.g., when present in an otherwise wildtype tRNA, recognition of the ribosome, e.g., acts as a recognition site for the ribosome to form a TREM-ribosome complex during translation. In an embodiment the THD has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring THD, e.g., a THD encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a THD, e.g., a THD encoded by a nucleic acid in Table 1, which fragment in embodiments has THD activity and in other embodiments does not have THD activity. In an embodiment the THD falls under the corresponding sequence of a consensus sequence provided in the “Consensus Sequence” section, or differs from the consensus sequence by no more than 1, 2, 5, or 10 positions; In an embodiment, the THD comprises residues -R48-R49-R50-R51-R52-R53-R54-R55-R56- R57-R58-R59-R60-R61-R62-R63-R64 of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids; In an embodiment, the THD comprises residues -R48-R49-R50-R51-R52-R53-R54-R55-R56- R57-R58-R59-R60-R61-R62-R63-R64 of Formula II ZZZ, wherein ZZZ indicates any of the twenty amino acids; Attorney Docket No.: F2099-7037WO In an embodiment, the THD comprises residues -R48-R49-R50-R51-R52-R53-R54-R55-R56- R57-R58-R59-R60-R61-R62-R63-R64 of Formula III ZZZ, wherein ZZZ indicates any of the twenty amino acids; (e’1) a linker comprising residue R72 of a consensus sequence provided in the “Consensus Sequence” section, e.g., a Linker 4 region; (f) under physiological conditions, it comprises a stem structure and one or a plurality of loop structures, e.g., 1, 2, or 3 loops. A loop can comprise a domain described herein, e.g., a domain selected from (a)-(e). A loop can comprise one or a plurality of domains. In an embodiment, a stem or loop structure has at least 75, 80, 85, 85, 90, 95, or 100% identity with a naturally occurring stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1. In an embodiment, the TREM can comprise a fragment or analog of a stem or loop structure, e.g., a stem or loop structure encoded by a nucleic acid in Table 1, which fragment in embodiments has activity of a stem or loop structure, and in other embodiments does not have activity of a stem or loop structure; (g) a tertiary structure, e.g., an L-shaped tertiary structure; (h) adaptor function, i.e., the TREM mediates acceptance of an amino acid, e.g., its cognate amino acid and transfer of the AA in the initiation or elongation of a polypeptide chain; (i) cognate adaptor function wherein the TREM mediates acceptance and incorporation of an amino acid (e.g., cognate amino acid) associated in nature with the anti-codon of the TREM to initiate or elongate a polypeptide chain; (j) non-cognate adaptor function, wherein the TREM mediates acceptance and incorporation of an amino acid (e.g., non-cognate amino acid) other than the amino acid associated in nature with the anti-codon of the TREM in the initiation or elongation of a polypeptide chain; (k) a regulatory function, e.g., an epigenetic function (e.g., gene silencing function or signaling pathway modulation function), cell fate modulation function, mRNA stability modulation function, protein stability modulation function, protein transduction modulation function, or protein compartmentalization function; (l) a structure which allows for ribosome binding; (m) a post-transcriptional modification, e.g., a naturally occurring post-trasncriptional modification; Attorney Docket No.: F2099-7037WO (n) the ability to inhibit a functional property of a tRNA, e.g., any of properties (h)-(k) possessed by a tRNA; (o) the ability to modulate cell fate; (p) the ability to modulate ribosome occupancy; (q) the ability to modulate protein translation; (r) the ability to modulate mRNA stability; (s) the ability to modulate protein folding and structure; (t) the ability to modulate protein transduction or compartmentalization; (u) the ability to modulate protein stability; or (v) the ability to modulate a signaling pathway, e.g., a cellular signaling pathway. In an embodiment, a TREM comprises a full-length tRNA molecule or a fragment thereof. In an embodiment, a TREM comprises the following properties: (a)-(e). In an embodiment, a TREM comprises the following properties: (a) and (c). In an embodiment, a TREM comprises the following properties: (a), (c) and (h). In an embodiment, a TREM comprises the following properties: (a), (c), (h) and (b). In an embodiment, a TREM comprises the following properties: (a), (c), (h) and (e). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (b) and (e). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (b), (e) and (g). In an embodiment, a TREM comprises the following properties: (a), (c), (h) and (m). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), and (g). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m) and (b). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m) and (e). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), (g), (b) and (e). In an embodiment, a TREM comprises the following properties: (a), (c), (h), (m), (g), (b), (e) and (q). In an embodiment, a TREM comprises: (i) an amino acid attachment domain that binds an amino acid (e.g., an AStD, as described in (a) herein; and Attorney Docket No.: F2099-7037WO (ii) an anticodon that binds a respective codon in an mRNA (e.g., an ACHD, as described in (c) herein). In an embodiment the TREM comprises a flexible RNA linker which provides for covalent linkage of (i) to (ii). In an embodiment, the TREM mediates protein translation. In an embodiment a TREM comprises a linker, e.g., an RNA linker, e.g., a flexible RNA linker, which provides for covalent linkage between a first and a second structure or domain. In an embodiment, an RNA linker comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 ribonucleotides. A TREM can comprise one or a plurality of linkers, e.g., in embodiments a TREM comprising (a), (b), (c), (d) and (e) can have a first linker between a first and second domain, and a second linker between a third domain and another domain. In an embodiment, the TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]- [L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2]. In an embodiment, a TREM comprises an RNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with, or which differs by no more than 1, 2, 3, 4, 5, 10, 15, 20, 25, or 30 ribonucleotides from, an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises an RNA sequence encoded by a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises an RNA sequence encoded by a DNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises a TREM domain, e.g., a domain described herein, comprising at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, or 99% identical with, or which differs by no more than 1, 2, 3, 4, 5, 10, or 15, ribonucleotides from, an RNA encoded by a DNA sequence listed in Table 1, or a fragment or a functional fragment thereof. In an embodiment, a TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by DNA sequence listed in Table 1, or a fragment or functional fragment thereof. In an embodiment, a TREM comprises a TREM domain, e.g., a domain described herein, comprising an RNA sequence encoded by DNA sequence at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98 or 99% identical with a DNA sequence listed in Table 1, or a fragment or functional fragment thereof. Attorney Docket No.: F2099-7037WO In an embodiment, a TREM is 76-90 nucleotides in length. In embodiments, a TREM or a fragment or functional fragment thereof is between 10-90 nucleotides, between 10-80 nucleotides, between 10-70 nucleotides, between 10-60 nucleotides, between 10-50 nucleotides, between 10-40 nucleotides, between 10-30 nucleotides, between 10-20 nucleotides, between 20- 90 nucleotides, between 20-80 nucleotides, 20-70 nucleotides, between 20-60 nucleotides, between 20-50 nucleotides, between 20-40 nucleotides, between 30-90 nucleotides, between 30- 80 nucleotides, between 30-70 nucleotides, between 30-60 nucleotides, or between 30-50 nucleotides. In an embodiment, a TREM is aminoacylated, e.g., charged, with an amino acid by an aminoacyl tRNA synthetase. In an embodiment, a TREM is not charged with an amino acid, e.g., an uncharged TREM (uTREM). In an embodiment, a TREM comprises less than a full length tRNA. In embodiments, a TREM can correspond to a naturally occurring fragment of a tRNA, or to a non-naturally occurring fragment. Exemplary fragments include: TREM halves (e.g., from a cleavage in the ACHD, e.g., in the anticodon sequence, e.g., 5’halves or 3’ halves); a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DHD or the ACHD); a 3’ fragment (e.g., a fragment comprising the 3’ end, e.g., from a cleavage in the THD); or an internal fragment (e.g., from a cleavage in one or more of the ACHD, DHD or THD). A “TREM core fragment,” as that term is used herein, refers to a portion of the sequence of Formula B: [L1] y-[ASt Domain1] x-[L2] y-[DH Domain]y-[L3] y-[ACH Domain]x-[VL Domain] y-[TH Domain] y-[L4] y-[ASt Domain2] x, wherein: x=1 and y=0 or 1. A “TREM fragment,” as used herein, refers to a portion of a TREM, wherein the TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2]. A “cognate adaptor function TREM,” as that term is used herein, refers to a TREM which mediates initiation or elongation with the AA (the cognate AA) associated in nature with the anti-codon of the TREM. “Decreased expression,” as that term is used herein, refers to a decrease in comparison to a reference, e.g., in the case where altered control region, or addition of an agent, results in a Attorney Docket No.: F2099-7037WO decreased expression of the subject product, it is decreased relative to an otherwise similar cell without the alteration or addition. An “exogenous nucleic acid,” as that term is used herein, refers to a nucleic acid sequence that is not present in or differs by at least one nucleotide from the closest sequence in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced. In an embodiment, an exogenous nucleic acid comprises a nucleic acid that encodes a TREM. An “exogenous TREM,” as that term is used herein, refers to a TREM that: (a) differs by at least one nucleotide or one post transcriptional modification from the closest sequence tRNA in a reference cell, e.g., a cell into which the exogenous nucleic acid is introduced; (b) has been introduced into a cell other than the cell in which it was transcribed; (c) is present in a cell other than one in which it naturally occurs; or (d) has an expression profile, e.g., level or distribution, that is non-wildtype, e.g., it is expressed at a higher level than wildtype. In an embodiment, the expression profile can be mediated by a change introduced into a nucleic acid that modulates expression or by addition of an agent that modulates expression of the RNA molecule. In an embodiment an exogenous TREM comprises 1, 2, 3 or 4 of properties (a)-(d). A “GMP-grade composition,” as that term is used herein, refers to a composition in compliance with current good manufacturing practice (cGMP) guidelines, or other similar requirements. In an embodiment, a GMP-grade composition can be used as a pharmaceutical product. As used herein, the terms “increasing” and “decreasing” refer to modulating that results in, respectively, greater or lesser amounts of function, expression, or activity of a particular metric relative to a reference. For example, subsequent to administration to a cell, tissue or subject of a TREM described herein, the amount of a marker of a metric (e.g., protein translation, mRNA stability, protein folding) as described herein may be increased or decreased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98%, 2X, 3X, 5X, 10X or more relative to the amount of the marker prior to administration or relative to the effect of a negative control agent. The metric may be measured subsequent to administration at a time that the administration has had the recited effect, e.g., at Attorney Docket No.: F2099-7037WO least 12 hours, 24 hours, one week, one month, 3 months, or 6 months, after a treatment has begun. “Increased expression,” as that term is used herein, refers to an increase in comparison to a reference, e.g., in the case where altered control region, or addition of an agent, results in an increased expression of the subject product, it is increased relative to an otherwise similar cell without the alteration or addition. A “non-cognate adaptor function TREM,” as that term is used herein, refers to a TREM which mediates initiation or elongation with an AA (a non-cognate AA) other than the AA associated in nature with the anti-codon of the TREM. In an embodiment, a non-cognate adaptor function TREM is also referred to as a mischarged TREM (mTREM). A “non-naturally occurring sequence,” as that term is used herein, refers to a sequence wherein an Adenine is replaced by a residue other than an analog of Adenine, a Cytosine is replaced by a residue other than an analog of Cytosine, a Guanine is replaced by a residue other than an analog of Guanine, and a Uracil is replaced by a residue other than an analog of Uracil. An analog refers to any possible derivative of the ribonucleotides, A, G, C or U. In an embodiment, a sequence having a derivative of any one of ribonucleotides A, G, C or U is a non- naturally occurring sequence. A “pharmaceutical TREM composition,” as that term is used herein, refers to a TREM composition that is suitable for pharmaceutical use. Typically, a pharmaceutical TREM composition comprises a pharmaceutical excipient. In an embodiment the TREM will be the only active ingredient in the pharmaceutical TREM composition. In embodiments the pharmaceutical TREM composition is free, substantially free, or has less than a pharmaceutically acceptable amount, of host cell proteins, DNA, e.g., host cell DNA, endotoxins, and bacteria. A “post-transcriptional processing,” as that term is used herein, with respect to a subject molecule, e.g., a TREM, RNA or tRNAs, refers to a covalent modification of the subject molecule. In an embodiment, the covalent modification occurs post-transcriptionally. In an embodiment, the covalent modification occurs co-transcriptionally. In an embodiment the modification is made in vivo, e.g., in a cell used to produce a TREM. In an embodiment the modification is made ex vivo, e.g., it is made on a TREM isolated or obtained from the cell which produced the TREM. In an embodiment, the post-transcriptional modification is selected from a post-transcriptional modification listed in Table 2. Attorney Docket No.: F2099-7037WO A “synthetic TREM,” as that term is used herein, refers to a TREM which was synthesized other than in or by a cell having an endogenous nucleic acid encoding the TREM, e.g., a synthetic TREM is synthetized by cell-free solid phase synthesis. A synthetic TREM can have the same, or a different, sequence, or tertiary structure, as a native tRNA. A “recombinant TREM,” as that term is used herein, refers to a TREM that was expressed in a cell modified by human intervention, having a modification that mediates the production of the TREM, e.g., the cell comprises an exogenous sequence encoding the TREM, or a modification that mediates expression, e.g., transcriptional expression or post-transcriptional modification, of the TREM. A recombinant TREM can have the same, or a different, sequence, set of post-transcriptional modifications, or tertiary structure, as a reference tRNA, e.g., a native tRNA. A “tRNA”, as that term is used herein, refers to a naturally occurring transfer ribonucleic acid in its native state. A “TREM composition,” as that term is used herein, refers to a composition comprising a plurality of TREMs, a plurality of TREM core fragments and/or a plurality of TREM fragments. A TREM composition can comprise one or more species of TREMs, TREM core fragments or TREM fragments. In an embodiment, the composition comprises only a single species of TREM, TREM core fragment or TREM fragment. In an embodiment, the TREM composition comprises a first TREM, TREM core fragment or TREM fragment species; and a second TREM, TREM core fragment or TREM fragment species. In an embodiment, the TREM composition comprises X TREM, TREM core fragment or TREM fragment species, wherein X=2, 3, 4, 5, 6, 7, 8, 9, or 10. In an embodiment, the TREM, TREM core fragment or TREM fragment has at least 70, 75, 80, 85, 90, or 95, or has 100%, identity with a sequence encoded by a nucleic acid in Table 1. A TREM composition can comprise one or more species of TREMs, TREM core fragments or TREM fragments. In an embodiment, the TREM composition is at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% dry weight TREMs (for a liquid composition dry weight refers to the weight after removal of substantially all liquid, e.g., after lyophilization). In an embodiment, the composition is a liquid. In an embodiment, the composition is dry, e.g., a lyophilized material. In an embodiment, the composition is a frozen composition. In an embodiment, the composition is sterile. In an embodiment, the composition comprises at least 0.5 g, 1.0 g, 5.0 g, 10 g, 15 g, 25 g, 50 g, 100 g, 200 g, 400 g, or 500 g (e.g., as determined by dry weight) of TREM. Attorney Docket No.: F2099-7037WO In an embodiment, at least X% of the TREMs in a TREM composition has a non- naturally occurring modification at a selected position, and X is 80, 90, 95, 96, 97, 98, 99, or 99.5. In an embodiment, at least X% of the TREMs in a TREM composition has a non- naturally occurring modification at a first position and a non-naturally occurring modification at a second position, and X, independently, is 80, 90, 95, 96, 97, 98, 99, or 99.5. In embodiments, the modification at the first and second position is the same. In embodiments, the modification at the first and second position are different. In embodiments, the nucleiotide at the first and second position is the same, e.g., both are adenine. In embodiments, the nucleiotide at the first and second position are different, e.g., one is adenine and one is thymine. In an embodiment, at least X% of the TREMs in a TREM composition has a non- naturally occurring modification at a first position and less than Y% have a non-naturally occurring modification at a second position, wherein X is 80, 90, 95, 96, 97, 98, 99, or 99.5 and Y is 20, 20, 5, 2, 1, 0.1, or 0.01. In embodiments, the nucleotide at the first and second position is the same, e.g., both are adenine. In embodiments the nucleotide at the first and second position are different, e.g., one is adenine and one is thymine. TREM, TREM core fragment and TREM fragment A “tRNA-based effector molecule” or “TREM” refers to an RNA molecule comprising one or more of the properties described herein. The TREMs described herein comprise a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide. A TREM comprising the LNA moiety or the 2’,5’-linked nucleotide can further comprise an additional non-naturally occurring modification, e.g., as provided in Table 5. In an embodiment, a TREM includes a TREM comprising a sequence of Formula A; a TREM core fragment comprising a sequence of Formula B; or a TREM fragment comprising a portion of a TREM which comprises a sequence of Formula A. In an embodiment, a TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]- [L2]-[DH Domain]-[L3]-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2]. In an embodiment, [VL Domain] is optional. In an embodiment, [L1] is optional. In an embodiment, a TREM core fragment comprises a sequence of Formula B: [L1] y- [ASt Domain1] x-[L2] y-[DH Domain]y-[L3] y-[ACH Domain]x-[VL Domain] y-[TH Domain] y- Attorney Docket No.: F2099-7037WO [L4] y-[ASt Domain2] x, wherein: x=1 and y=0 or 1. In an embodiment, y=0. In an embodiment, y=1.; In an embodiment, a TREM fragment comprises a portion of a TREM, wherein the TREM comprises a sequence of Formula A: [L1]-[ASt Domain1]-[L2]-[DH Domain]-[L3]- [ACH Domain] -[VL Domain]-[TH Domain]-[L4]-[ASt Domain2], and wherein the TREM fragment comprises: one, two, three or all or any combination of the following: a TREM half (e.g., from a cleavage in the ACH Domain, e.g., in the anticodon sequence, e.g., a 5’half or a 3’ half); a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DH Domain or the ACH Domain); a 3’ fragment (e.g., a fragment comprising the 3’ end, e.g., from a cleavage in the TH Domain); or an internal fragment (e.g., from a cleavage in any one of the ACH Domain, DH Domain or TH Domain). Exemplary TREM fragments include TREM halves (e.g., from a cleavage in the ACHD, e.g., 5’TREM halves or 3’ TREM halves), a 5’ fragment (e.g., a fragment comprising the 5’ end, e.g., from a cleavage in a DHD or the ACHD), a 3’ fragment (e.g., a fragment comprising the 3’ end of a TREM, e.g., from a cleavage in the THD), or an internal fragment (e.g., from a cleavage in one or more of the ACHD, DHD or THD). In an embodiment, a TREM, a TREM core fragment or a TREM fragment can be charged with an amino acid (e.g., a cognate amino acid); charged with a non-cognate amino acid (e.g., a mischarged TREM (mTREM)); or not charged with an amino acid (e.g., an uncharged TREM (uTREM)). In an embodiment, a TREM, a TREM core fragment or a TREM fragment can be charged with an amino acid selected from alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine. In some embodiments, a non-extended anticodon is an anticodon of no more than three nucleotides. In an embodiment, a non-extended codon pairs with no more than three codon nucleotides on a nucleic acid being translated. In an embodiment, the TREM, TREM core fragment or TREM fragment is a cognate TREM. In an embodiment, the TREM, TREM core fragment or TREM fragment is a non- cognate TREM. In an embodiment, the TREM, TREM core fragment or TREM fragment recognizes a codon provided in Table 2 or Table 3. Table 2: List of codons Attorney Docket No.: F2099-7037WO AAA AAC
Figure imgf000030_0001
Attorney Docket No.: F2099-7037WO GGA GGC
Figure imgf000031_0001
Table 3: Amino acids and corresponding codons Amino Acid mRNA codons
Figure imgf000031_0002
Attorney Docket No.: F2099-7037WO Phenylalanine UUU, UUC Proline CCU, CCC, CCA, CCG
Figure imgf000032_0001
, p leic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1- 451 disclosed in Table 1. In an embodiment, a TREM comprises an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM comprises an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM, a TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM, a TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM, a TREM core fragment, or TREM fragment comprises at least 5, 10, 15, 20, 25, or 30 consecutive nucleotides of an RNA sequence encoded by a DNA sequence at least 60%, 65%, 70%, 75%, 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM core fragment or a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, Attorney Docket No.: F2099-7037WO 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM core fragment or a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM core fragment or a TREM fragment comprises at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of an RNA sequence encoded by a DNA sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM core fragment or a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence disclosed in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM core fragment or a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to an RNA sequence encoded by a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM core fragment or a TREM fragment comprises at least 5 ribonucleotides (nt), 10 nt, 15 nt, 20 nt, 25 nt, 30 nt, 35 nt, 40 nt, 45 nt, 50 nt, 55 nt or 60 nt (but less than the full length) of an RNA sequence encoded by a DNA sequence with at least 80%, 82%, 85%, 87%, 88%, 90%, 92%, 95%, 96%, 97%, 98%, 99% or 100% identity to a DNA sequence provided in Table 1, e.g., any one of SEQ ID NOs: 1-451 disclosed in Table 1. In an embodiment, a TREM core fragment or a TREM fragment comprises a sequence of a length of between 10-90 ribonucleotides (rnt), between 10-80 rnt, between 10-70 rnt, between 10-60 rnt, between 10-50 rnt, between 10-40 rnt, between 10-30 rnt, between 10-20 rnt, between 20-90 rnt, between 20-80 rnt, 20-70 rnt, between 20-60 rnt, between 20-50 rnt, between 20-40 rnt, between 30-90 rnt, between 30-80 rnt, between 30-70 rnt, between 30-60 rnt, or between 30- 50 rnt. Table 1: List of tRNA Sequences Attorney Docket No.: F2099-7037WO SEQ ID tRNA name tRNA sequence NO T G T T T T T G T G T G A T G
Figure imgf000034_0001
Attorney Docket No.: F2099-7037WO Ala_AGC_chr2:27274 GGGGGATTAGCTCAAATGGTAGAGCGCTCGC 082-27274154 (+) TTAGCATGCGAGAGGTAGCGGGATCGATGCC T G T G G T G T G T G T G T G T A T G T C T T
Figure imgf000035_0001
Attorney Docket No.: F2099-7037WO Arg_CCG_chr6:28710 GGCCGCGTGGCCTAATGGATAAGGCGTCTGAT 729-28710801 (-) TCCGGATCAGAAGATTGAGGGTTCGAGTCCCT C C C C A C A C A T C C C C C G G C G
Figure imgf000036_0001
Attorney Docket No.: F2099-7037WO Arg_TCT_chr9:13110 GGCTCTGTGGCGCAATGGATAGCGCATTGGAC 2355-131102445 (-) TTCTAGCTGAGCCTAGTGTGGTCATTCAAAGG C G C C G A G A A G A G A G A G A G A G A A G A
Figure imgf000037_0001
Attorney Docket No.: F2099-7037WO Asn_GTT_chr1:14597 GTCTCTGTGGCGCAATCGGTTAGCGCGTTCGG 9034-145979107 (-) CTGTTAACTGAAAGGTTAGTGGTTCGAGCCCA G G G T G T T G G G G G T G
Figure imgf000038_0001
Attorney Docket No.: F2099-7037WO Cys_GCA_chr4:12443 GGGGGTATAGCTCAGTGGTAGAGCATTTGACT 0005-124430076 (-) GCAGATCAAGAGGTCCCCGGTTCAAATCCGG T T T T T G T G T G G G C G C G C G
Figure imgf000039_0001
Attorney Docket No.: F2099-7037WO Gln_CTG_chr1:14773 GGTTCCATGGTGTAATGGTAAGCACTCTGGAC 7382-147737453 (-) TCTGAATCCAGCGATCCGAGTTCGAGTCTCGG C G C G C C G C G C G C C G C G G G C C C
Figure imgf000040_0001
Attorney Docket No.: F2099-7037WO Gly_CCC_chr2:70476 GCGCCGCTGGTGTAGTGGTATCATGCAAGATT 123-70476193 (-) CCCATTCTTGCGACCCGGGTTCGATTCCCGGG C T C T C C C G C G C G C C G G G C G C
Figure imgf000041_0001
Attorney Docket No.: F2099-7037WO Ile_AAT_chr6:272429 GGCTGGTTAGCTCAGTTGGTTAGAGCGTGGTG 90-27243063 (-) CTAATAACGCCAAGGTCGCGGGTTCGATCCCC G C G C G C G C G C G C G C A A A G A G A G T T
Figure imgf000042_0001
Attorney Docket No.: F2099-7037WO Leu_AAG_chr6:2895 GGTAGCGTGGCCGAGCGGTCTAAGGCGCTGG 6779-28956860 (+) ATTAAGGCTCCAGTCTCTTCGGGGGCGTGGGT T T C C T C C C C C C G G G G G
Figure imgf000043_0001
Attorney Docket No.: F2099-7037WO Leu_TAA_chr11:5931 ACCAGAATGGCCGAGTGGTTAAGGCGTTGGA 9228-59319310 (+) CTTAAGATCCAATGGATTCATATCCGCGTGGG G T T C
Figure imgf000044_0001
Attorney Docket No.: F2099-7037WO Lys_TTT_chr16:7351 GCCTGGATAGCTCAGTTGGTAGAGCATCAGAC 2216-73512288 (-) TTTTAATCTGAGGGTCCAGGGTTCAAGTCCCT C C C C A C A C A C A C A T A
Figure imgf000045_0001
Attorney Docket No.: F2099-7037WO Phe_GAA_chr6:28758 GCCGAAATAGCTCAGTTGGGAGAGCGTTAGA 499-28758571 (-) CTGAAGATCTAAAGGTCCCTGGTTCGATCCCG G G C T G G T G T T
Figure imgf000046_0001
Attorney Docket No.: F2099-7037WO Ser_AGA_chr6:26327 GTAGTCGTGGCCGAGTGGTTAAGGCGATGGA 817-26327898 (+) CTAGAAATCCATTGGGGTCTCCCCGCGCAGGT T T C T C T C T T T T
Figure imgf000047_0001
Attorney Docket No.: F2099-7037WO Ser_TGA_chr6:26312 GTAGTCGTGGCCGAGTGGTTAAGGCGATGGA 824-26312905 (-) CTTGAAATCCATTGGGGTCTCCCCGCGCAGGT T T C T C T C T C T C T C C T C C C C C G
Figure imgf000048_0001
Attorney Docket No.: F2099-7037WO Thr_TGT_chr14:2108 GGCTCCATAGCTCAGGGGTTAGAGCGCTGGTC 1949-21082021 (-) TTGTAAACCAGGGGTCGCGAGTTCAATTCTCG C G C G C C T T T T T T T T T T C C C C C C
Figure imgf000049_0001
Attorney Docket No.: F2099-7037WO Tyr_GTA_chr8:67026 CCTTCGATAGCTCAGCTGGTAGAGCGGAGGA 223-67026311 (+) CTGTAGGCGCGCGCCCGTGGCCATCCTTAGGT T T T C T C G T T G
Figure imgf000050_0001
Attorney Docket No.: F2099-7037WO Val_CAC_chr6:27248 GCTTCTGTAGTGTAGTGGTTATCACGTTCGCC 049-27248121 (-) TCACACGCGAAAGGTCCCCGGTTCGAAACCG G G A A A C C T C C C G G
Figure imgf000051_0001
Attorney Docket No.: F2099-7037WO Asn_GTT_chr1:17216 TGTCTCTGTGGCGCAATCGGTTAGCGCGTTCG 171-17216245 (+) GCTGTTAACCGAAAGATTGGTGGTTCGAGCCC A G G C G C G A G C C G A G A T T T T G
Figure imgf000052_0001
Attorney Docket No.: F2099-7037WO Met_CAT_chr1:15364 TAGCAGAGTGGCGCAGCGGAAGCGTGCTGGG 3725-153643797 (+) CCCATAACCCAGAGGTCGATGGATCGAAACC G C G C G C G C C C G
Figure imgf000053_0001
Attorney Docket No.: F2099-7037WO Leu_CAA_chr1:24916 TGTCAGGATGGCCGAGTGGTCTAAGGCGCCA 8053-249168159 (+) GACTCAAGGTAAGCACCTTGCCTGCGGGCTTT C G T T G T G
Figure imgf000054_0001
Attorney Docket No.: F2099-7037WO Thr_TGT_chr5:18061 GGCTCCATAGCTCAGGGGTTAGAGCACTGGTC 8686-180618758 (-) TTGTAAACCAGGGTCGCGAGTTCAAATCTCGC T G G C G C G T C C G C
Figure imgf000055_0001
Attorney Docket No.: F2099-7037WO Leu_CAG_chr6:26521 CGTCAGGATGGCCGAGCGGTCTAAGGCGCTG 435-26521518 (+) CGTTCAGGTCGCAGTCTCCCCTGGAGGCGTGG T C C T C A T T C A G C
Figure imgf000056_0001
Attorney Docket No.: F2099-7037WO Val_CAC_chr6:27248 GCTTCTGTAGTGTAGTGGTTATCACGTTCGCC 048-27248121 (-) TCACACGCGAAAGGTCCCCGGTTCGAAACCG C T A G G T C C G C G T G T G C T T T C
Figure imgf000057_0001
Attorney Docket No.: F2099-7037WO Leu_CAA_chr6:28908 TGTCAGGATGGCCGAGTGGTCTAAGGCGCCA 829-28908934 (+) GACTCAAGCTTGGCTTCCTCGTGTTGAGGATT C C G T T C A G C G T G T
Figure imgf000058_0001
Attorney Docket No.: F2099-7037WO Met_CAT_chr8:12416 GCCTCGTTAGCGCAGTAGGTAGCGCGTCAGTC 9469-124169542 (-) TCATAATCTGAAGGTCGTGAGTTCGATCCTCA C G G A G A A A T G G G G G
Figure imgf000059_0001
Attorney Docket No.: F2099-7037WO Asp_GTC_chr12:9889 CTCCTCGTTAGTATAGTGGTTAGTATCCCCGC 7280-98897352 (+) CTGTCACGCGGGAGACCGGGGTTCAATTCCCC C T G G G A G C G G C G T T A T C A G
Figure imgf000060_0001
Attorney Docket No.: F2099-7037WO Lys_CTT_chr14:5870 GCCCGGCTAGCTCAGTCGGTAGAGCATGGGA 6612-58706685 (-) CTCTTAATCCCAGGGTCGTGGGTTCGAGCCCC T C G G A C G C C T C C C C
Figure imgf000061_0001
Attorney Docket No.: F2099-7037WO Leu_AAG_chr16:223 GGGTAGCGTGGCCGAGCGGTCTAAGGCGCTG 08460-22308542 (+) GATTAAGGCTCCAGTCTCTTCGGGGGCGTGGG G G C A T A T G C T T C C G G C T T C G
Figure imgf000062_0001
Attorney Docket No.: F2099-7037WO Pro_CGG_chr17:8126 GGCTCGTTGGTCTAGGGGTATGATTCTCGCTT 150-8126222 (-) CGGGTGCGAGAGGTCCCGGGTTCAAATCCCG T C T C G C G T T A C C C C C C
Figure imgf000063_0001
Attorney Docket No.: F2099-7037WO 448 Val_CAC_chr19:4724 GTTTCCGTAGTGTAGCGGTTATCACATTCGCC 646-4724719 (-) TCACACGCGAAAGGTCCCCGGTTCGATCCCGG G C A T
Figure imgf000064_0001
Non-naturally occurring modification A TREM, a TREM core fragment or a TREM fragment described herein comprises a non-naturally occurring modification, e.g., a modification described in Table 5. A non-naturally occurring modification can be made according to methods known in the art. Exemplary methods of making non-naturally occurring modifications are provided in Example 1. In an embodiment, a non-naturally occurring modification is a modification that a cell, e.g., a human cell, does not make on an endogenous tRNA. In an embodiment, a non-naturally occurring modification is a modification that a cell, e.g., a human cell, can make on an endogenous tRNA, but wherein such modification is in a location in which it does not occur on a native tRNA. In an embodiment, the non-naturally occurring modification is in a domain, linker or arm which does not have such modification in nature. In an embodiment, the non-naturally occurring modification is at a position within a domain, linker or arm, which does not have such modification in nature. In an embodiment, the non-naturally occurring modification is on a nucleotide which does not have such modification in nature. In an embodiment, the non-naturally occurring modification is on a nucleotide at a position within a domain, linker or arm, which does not have such modification in nature.
Attorney Docket No.: F2099-7037WO In an embodiment, a TREM, a TREM core fragment or a TREM fragment described herein comprises a modification provided in Table 5, or a combination thereof. The modifications provided in Table 5 are non-naturally occurring or occur naturally in RNAs, and are used herein on a synthetic TREM, a TREM core fragment or a TREM fragment at a position that does not occur in nature. Table 5: Exemplary modifications Chemical Modification (S)-constrained ethyl (cEt) 5-(methoxycarbonyl-methyl)uracil
Figure imgf000065_0001
Attorney Docket No.: F2099-7037WO 1-(3-Amino-3-carboxypropyl)pseudo-uridine 5-carbamoylmethyl-2’-O-methyluridine 1-(3-Amino-propyl)pseudouridine 5-carbamoylmethyluridine
Figure imgf000066_0001
Attorney Docket No.: F2099-7037WO 1-{3-[2-(2-Aminoethoxy)-ethoxy]- propionvl}pseudouridine 5-methyluridine -
Figure imgf000067_0001
Attorney Docket No.: F2099-7037WO -Methyl-6-hydroxyamino-pseudo-uridine 6-Cyano-pseudo-uridine -Methyl-6-trifluoromethoxy-pseudo-uridine 6-Dimethylamino-pseudo-uridine
Figure imgf000068_0001
Attorney Docket No.: F2099-7037WO ’-alpha-Trifluoromethyluridine 7-(guanidiniumalkylhydroxy)-1-(aza)-2-(thio)-3- (aza)-phenoxazin-1-yl )- n- l
Figure imgf000069_0001
Attorney Docket No.: F2099-7037WO ’-Deoxy-2’-beta-aminocytidine 8-(hydroxyl)guanine ’-Deoxy-2’-beta-bromoadenosine 8-(thioalkyl)adenine - -
Figure imgf000070_0001
Attorney Docket No.: F2099-7037WO ’-O-Methyl-5-(1-propynyl)cytidine -CH2-NH-CH2- ’-O-Methyl-5-(1-propynyl)uridine chiral phosphonates
Figure imgf000071_0001
Attorney Docket No.: F2099-7037WO -Azidoadenosine l -Methyl-6-iodo-pseudo-uridine ’-beta-Trifluoromethyladenosine l -Methyl-6-methylamino-pseudo-uridine -
Figure imgf000072_0001
Attorney Docket No.: F2099-7037WO -methylthio-N6-threonyl carbamoyladenosine methylphosphonates ’-O-methoxyethyl (MOE) methylwyosine
Figure imgf000073_0001
Attorney Docket No.: F2099-7037WO -(methyl)isocarbostyrilyl N6-isopentenyladenosine ,2’-0-dimethyluridine N6-methyl-2-amino-purine e
Figure imgf000074_0001
Attorney Docket No.: F2099-7037WO -methoxy-pseudoisocytidine PNA -methoxy-pseudouridine propynyl-7-(aza)indolyl
Figure imgf000075_0001
Attorney Docket No.: F2099-7037WO 5-(alkynyl)cytosine sulfamate backbones 5-(alkynyl)uracil sulfide sulfoxide and sulfone backbones
Figure imgf000076_0001
Chemically Modified TREMs The present disclosure features TREMs comprising a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide, as described herein. In some embodiments, the TREM may comprise both LNA moieties and 2’,5’-linked nucleotides, optionally further comprising at least one additional non-naturally occurring modification In some embodiments, the TREM comprising the LNA moiety or the 2’,5’-linked nucleotide further comprises at least one additional non-naturally occurring modification (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications). Locked Nucleic Acids Described herein are TREMs comprising a nucleotide comprising a locked nucleic acid moiety. The term “locked nucleic acid (LNA) moiety,” as used herein, refers to a nucleotide comprising a modified nucleotide sugar, in which the 2’-carbon is linked to the 4’-carbon atom through a cyclic structure. In an embodiment, the LNA moiety comprises a heterocyclyl ring or cycloalkyl ring between the 2’-position and the 4’-position of the nucleotide sugar within the Attorney Docket No.: F2099-7037WO nucleotide. In an embodiment, the TREM is a sequence of Formula (I), wherein the heterocyclyl ring comprises an oxygen atom, nitrogen atom, sulfur atom, selenium atom. In some embodiments, the heterocyclyl ring or cycloalkyl ring is a five-membered ring, six-membered ring, seven-membered ring, or eight-membered ring. In one aspect, the present disclosure features a TREM containing a nucleotide comprising the LNA moiety having the structure of Formula (II): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a (RB); each of R1, R2, and R3 is independently hydrogen,
Figure imgf000077_0001
C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, - ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and X is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, - S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, - N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or - Attorney Docket No.: F2099-7037WO ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; provided that no more than two of L, M, Z, and Z may be -
Figure imgf000078_0001
, - -, x, - Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. In some embodiments, B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). In some embodiments, B is a purine-based nucleobase. In some embodiments, B is a pyrimidine-based nucleobase. In some embodiments, B is adenine. In some embodiments, B is cytosine. In some embodiments, B is guanine. In some embodiments, B is uracil. In some embodiments, B is a non-naturally occurring nucleobase. In some embodiments, B comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. In some embodiments, the chemical modification is a nucleotide sugar modification. In some embodiments, the chemical modification is a 2’OMe modification. In some embodiments, the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification. In some embodiments, the chemical modification is a 2’MOE modification. In some embodiments, the chemical modification is a 2’-deoxy modification. In some embodiments, the chemical modification is present in the internucleotide region (e.g., a backbone modification). In some embodiments, the chemical modification is a phosphorothioate modification. In some embodiments, B does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, Y is selected from CH2, S, or O. In an embodiment, Y is CH2. In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R1 is hydrogen. In an embodiment, R1 is C1-C6 alkyl. In an embodiment, R1 is C1- C6 heteroalkyl. In an embodiment, R1 is C1-C6 haloalkyl. In an embodiment, R1 is halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an Attorney Docket No.: F2099-7037WO embodiment, R2 is hydrogen. In an embodiment, R2 is C1-C6 alkyl. In an embodiment, R2 is C1- C6 heteroalkyl. In an embodiment, R2 is C1-C6 haloalkyl. In an embodiment, R2 is halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R3 is hydrogen. In an embodiment, R3 is C1-C6 alkyl. In an embodiment, R3 is C1- C6 heteroalkyl. In an embodiment, R3 is C1-C6 haloalkyl. In an embodiment, R3 is halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently hydrogen. In an embodiment, each of R4a and R4b is independently C1-C6 alkyl. In an embodiment, each of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, each of R4a and R4b is independently halo. In an embodiment, one of R4a and R4b is independently hydrogen. In an embodiment, one of R4a and R4b is independently C1-C6 alkyl. In an embodiment, one of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, one of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, one of R4a and R4b is independently halo. In some embodiments, one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(RB)-. In some embodiments, one of L and X is O. In some embodiments, one of L and X is N(RB)-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(RB). In some embodiments, X is S. In some embodiments, X is Se. In some embodiments, the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-a): Y Attorney Docket No.: F2099-7037WO is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; each of Ria, Rib, Rii, and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. In some embodiments, Y is selected from CH2, S, or O. In an embodiment, Y is CH2. In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R1 is hydrogen. In an embodiment, R1 is C1-C6 alkyl. In an embodiment, R1 is C 1- C6 heteroalkyl. In an embodiment, R1 is C1-C6 haloalkyl. In an embodiment, R1 is halo. Attorney Docket No.: F2099-7037WO In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R2 is hydrogen. In an embodiment, R2 is C1-C6 alkyl. In an embodiment, R2 is C1- C6 heteroalkyl. In an embodiment, R2 is C1-C6 haloalkyl. In an embodiment, R2 is halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R3 is hydrogen. In an embodiment, R3 is C1-C6 alkyl. In an embodiment, R3 is C1- C6 heteroalkyl. In an embodiment, R3 is C1-C6 haloalkyl. In an embodiment, R3 is halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently hydrogen. In an embodiment, each of R4a and R4b is independently C1-C6 alkyl. In an embodiment, each of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, each of R4a and R4b is independently halo. In an embodiment, one of R4a and R4b is independently hydrogen. In an embodiment, one of R4a and R4b is independently C1-C6 alkyl. In an embodiment, one of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, one of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, one of R4a and R4b is independently halo. In some embodiments, one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(RB)-. In some embodiments, one of L and X is O. In some embodiments, one of L and X is N(RB)-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(RB). In some embodiments, X is S. In some embodiments, X is Se. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of Ria and Rib is independently hydrogen. In an embodiment, each of Ria and Rib is independently C1-C6 alkyl. In an embodiment, each of Ria and Rib is independently C1-C6 heteroalkyl. In an embodiment, each of Ria and Rib is independently C 1 -C 6 haloalkyl. In an embodiment, one of Ria and Rib is independently hydrogen. In an embodiment, one of Ria and Rib is independently C1-C6 alkyl. In Attorney Docket No.: F2099-7037WO an embodiment, one of Ria and Rib is independently C1-C6 heteroalkyl. In an embodiment, one of Ria and Rib is independently C1-C6 haloalkyl. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently hydrogen. In an embodiment, each of Rii and Riii is independently C1-C6 alkyl. In an embodiment, each of Rii and Riii is independently C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, each of Rii and Riii is independently halo. In an embodiment, one of Rii and Riii is independently hydrogen. In an embodiment, one of Rii and Riii is independently C1-C6 alkyl. In an embodiment, one of Rii and Riii is independently C1-C6 heteroalkyl. In an embodiment, one of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, one of Rii and Riii is independently halo. In some embodiments, the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-b): a salt, stereoisomer, tautomer, or hydrate thereof,
Figure imgf000082_0001
R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or - N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, - Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, - Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1- Attorney Docket No.: F2099-7037WO C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; each of Ria, Rib, Rii, and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. In some embodiments, Y is selected from CH2, S, or O. In an embodiment, Y is CH2. In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R1 is hydrogen. In an embodiment, R1 is C1-C6 alkyl. In an embodiment, R1 is C1- C6 heteroalkyl. In an embodiment, R1 is C1-C6 haloalkyl. In an embodiment, R1 is halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R2 is hydrogen. In an embodiment, R2 is C1-C6 alkyl. In an embodiment, R2 is C1- C6 heteroalkyl. In an embodiment, R2 is C1-C6 haloalkyl. In an embodiment, R2 is halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R3 is hydrogen. In an embodiment, R3 is C1-C6 alkyl. In an embodiment, R3 is C1- C6 heteroalkyl. In an embodiment, R3 is C1-C6 haloalkyl. In an embodiment, R3 is halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R4a and R4b is Attorney Docket No.: F2099-7037WO independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently hydrogen. In an embodiment, each of R4a and R4b is independently C1-C6 alkyl. In an embodiment, each of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, each of R4a and R4b is independently halo. In an embodiment, one of R4a and R4b is independently hydrogen. In an embodiment, one of R4a and R4b is independently C1-C6 alkyl. In an embodiment, one of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, one of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, one of R4a and R4b is independently halo. In some embodiments, one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(RB)-. In some embodiments, one of L and X is O. In some embodiments, one of L and X is N(RB)-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(RB). In some embodiments, X is S. In some embodiments, X is Se. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of Ria and Rib is independently hydrogen. In an embodiment, each of Ria and Rib is independently C1-C6 alkyl. In an embodiment, each of Ria and Rib is independently C1-C6 heteroalkyl. In an embodiment, each of Ria and Rib is independently C1-C6 haloalkyl. In an embodiment, one of Ria and Rib is independently hydrogen. In an embodiment, one of Ria and Rib is independently C1-C6 alkyl. In an embodiment, one of Ria and Rib is independently C1-C6 heteroalkyl. In an embodiment, one of Ria and Rib is independently C1-C6 haloalkyl. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently hydrogen. In an embodiment, each of Rii and Riii is independently C1-C6 alkyl. In an embodiment, each of Rii and Riii is independently C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, each of Rii and Riii is independently halo. In an embodiment, one of Rii and Riii is independently hydrogen. In an embodiment, one of Rii and Riii is independently C1-C6 alkyl. In an embodiment, one of Rii and Attorney Docket No.: F2099-7037WO Riii is independently C1-C6 heteroalkyl. In an embodiment, one of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, one of Rii and Riii is independently halo. In some embodiments, the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-c): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y is and R3 is independently hydrogen, C1-C6 alkyl, C2-C6
Figure imgf000085_0001
alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; each of Ria, Rib, Rii, and Riii is independently Attorney Docket No.: F2099-7037WO hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. In some embodiments, Y is selected from CH2, S, or O. In an embodiment, Y is CH2. In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R1 is hydrogen. In an embodiment, R1 is C1-C6 alkyl. In an embodiment, R1 is C1- C6 heteroalkyl. In an embodiment, R1 is C1-C6 haloalkyl. In an embodiment, R1 is halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R2 is hydrogen. In an embodiment, R2 is C1-C6 alkyl. In an embodiment, R2 is C1- C6 heteroalkyl. In an embodiment, R2 is C1-C6 haloalkyl. In an embodiment, R2 is halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R3 is hydrogen. In an embodiment, R3 is C1-C6 alkyl. In an embodiment, R3 is C1- C6 heteroalkyl. In an embodiment, R3 is C1-C6 haloalkyl. In an embodiment, R3 is halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently hydrogen. In an embodiment, each of R4a and R4b is independently C1-C6 alkyl. In an embodiment, each of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, each of R4a and R4b is independently halo. In an embodiment, one of R4a and R4b is independently hydrogen. In an embodiment, one of R4a and R4b is independently C1-C6 alkyl. In an embodiment, one of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, one of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, one of R4a and R4b is independently halo. Attorney Docket No.: F2099-7037WO In some embodiments, one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(RB)-. In some embodiments, one of L and X is O. In some embodiments, one of L and X is N(RB)-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(RB). In some embodiments, X is S. In some embodiments, X is Se. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of Ria and Rib is independently hydrogen. In an embodiment, each of Ria and Rib is independently C1-C6 alkyl. In an embodiment, each of Ria and Rib is independently C1-C6 heteroalkyl. In an embodiment, each of Ria and Rib is independently C1-C6 haloalkyl. In an embodiment, one of Ria and Rib is independently hydrogen. In an embodiment, one of Ria and Rib is independently C1-C6 alkyl. In an embodiment, one of Ria and Rib is independently C1-C6 heteroalkyl. In an embodiment, one of Ria and Rib is independently C1-C6 haloalkyl. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently hydrogen. In an embodiment, each of Rii and Riii is independently C1-C6 alkyl. In an embodiment, each of Rii and Riii is independently C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, each of Rii and Riii is independently halo. In an embodiment, one of Rii and Riii is independently hydrogen. In an embodiment, one of Rii and Riii is independently C1-C6 alkyl. In an embodiment, one of Rii and Riii is independently C1-C6 heteroalkyl. In an embodiment, one of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, one of Rii and Riii is independently halo. In some embodiments, the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-d): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6
Figure imgf000088_0001
C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; each of Ri, Rii, and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or
Figure imgf000088_0002
of the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, Attorney Docket No.: F2099-7037WO S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. In some embodiments, Y is selected from CH2, S, or O. In an embodiment, Y is CH2. In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R1 is hydrogen. In an embodiment, R1 is C1-C6 alkyl. In an embodiment, R1 is C1- C6 heteroalkyl. In an embodiment, R1 is C1-C6 haloalkyl. In an embodiment, R1 is halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R2 is hydrogen. In an embodiment, R2 is C1-C6 alkyl. In an embodiment, R2 is C1- C6 heteroalkyl. In an embodiment, R2 is C1-C6 haloalkyl. In an embodiment, R2 is halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R3 is hydrogen. In an embodiment, R3 is C1-C6 alkyl. In an embodiment, R3 is C1- C6 heteroalkyl. In an embodiment, R3 is C1-C6 haloalkyl. In an embodiment, R3 is halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently hydrogen. In an embodiment, each of R4a and R4b is independently C1-C6 alkyl. In an embodiment, each of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, each of R4a and R4b is independently halo. In an embodiment, one of R4a and R4b is independently hydrogen. In an embodiment, one of R4a and R4b is independently C1-C6 alkyl. In an embodiment, one of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, one of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, one of R4a and R4b is independently halo. In some embodiments, one of L, M, Z, and X is O. In some embodiments, one of L, M, Z, and X is N(RB)-. In some embodiments, one of L and X is O. In some embodiments, one of L and X is N(RB)-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE) y -. In some embodiments, X is selected from C 1 -C 6 alkylene, O, N(RB), S, and Se. In Attorney Docket No.: F2099-7037WO some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(RB). In some embodiments, X is S. In some embodiments, X is Se. In some embodiments, Ri is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, Ri is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, Ri is hydrogen. In an embodiment, Ri is C1-C6 alkyl. In an embodiment, Ri is C1-C6 heteroalkyl. In an embodiment, Ri is C1-C6 haloalkyl. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently hydrogen. In an embodiment, each of Rii and Riii is independently C1-C6 alkyl. In an embodiment, each of Rii and Riii is independently C1-C6 heteroalkyl. In an embodiment, each of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, each of Rii and Riii is independently halo. In an embodiment, one of Rii and Riii is independently hydrogen. In an embodiment, one of Rii and Riii is independently C1-C6 alkyl. In an embodiment, one of Rii and Riii is independently C1-C6 heteroalkyl. In an embodiment, one of Rii and Riii is independently C1-C6 haloalkyl. In an embodiment, one of Rii and Riii is independently halo. In some embodiments, the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-e): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a
Figure imgf000090_0001
; each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, - ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein Attorney Docket No.: F2099-7037WO each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; n is 1, 2, 3, 4, 5, or 6; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. In some embodiments, Y is selected from CH2, S, or O. In an embodiment, Y is CH2. In an embodiment, Y is S. In an embodiment, Y is O. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R1 is hydrogen. In an embodiment, R1 is C1-C6 alkyl. In an embodiment, R1 is C1- C6 heteroalkyl. In an embodiment, R1 is C1-C6 haloalkyl. In an embodiment, R1 is halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R2 is hydrogen. In an embodiment, R2 is C1-C6 alkyl. In an embodiment, R2 is C1- C6 heteroalkyl. In an embodiment, R2 is C1-C6 haloalkyl. In an embodiment, R2 is halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, R3 is hydrogen. In an embodiment, R3 is C1-C6 alkyl. In an embodiment, R3 is C1- C6 heteroalkyl. In an embodiment, R3 is C1-C6 haloalkyl. In an embodiment, R3 is halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently hydrogen. In an embodiment, each of R4a and R4b is independently C1-C6 alkyl. Attorney Docket No.: F2099-7037WO In an embodiment, each of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, each of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, each of R4a and R4b is independently halo. In an embodiment, one of R4a and R4b is independently hydrogen. In an embodiment, one of R4a and R4b is independently C1-C6 alkyl. In an embodiment, one of R4a and R4b is independently C1-C6 heteroalkyl. In an embodiment, one of R4a and R4b is independently C1-C6 haloalkyl. In an embodiment, one of R5a and R5b is independently halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R5a and R5b is independently hydrogen. In an embodiment, each of R5a and R5b is independently C1-C6 alkyl. In an embodiment, each of R5a and R5b is independently C1-C6 heteroalkyl. In an embodiment, each of R5a and R5b is independently C1-C6 haloalkyl. In an embodiment, each of R5a and R5b is independently halo. In an embodiment, one of R5a and R5b is independently hydrogen. In an embodiment, one of R5a and R5b is independently C1-C6 alkyl. In an embodiment, one of R5a and R5b is independently C1-C6 heteroalkyl. In an embodiment, one of R5a and R5b is independently C1-C6 haloalkyl. In an embodiment, one of R5a and R5b is independently halo. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and - Se(RE)y-. In some embodiments, X is selected from C1-C6 alkylene, O, N(RB), S, and Se. In some embodiments, X is O. In some embodiments, X is C1-C6 alkylene. In some embodiments, X is N(RB). In some embodiments, X is S. In some embodiments, X is Se. In some embodiments, n is selected from 1, 2, or 3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-e-i) or (II-e-ii): e-ii), or a salt, stereoisomer,
Figure imgf000092_0001
Attorney Docket No.: F2099-7037WO In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-f): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl,
Figure imgf000093_0001
C2-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1- C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1- C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or - ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; n is 1, 2, 3, 4, 5, or 6; and “ ” refers to an attachment point to the rest of the TREM sequence, another
Figure imgf000093_0002
of the TREM. In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-f-i) or (II-f-ii): Attorney Docket No.: F2099-7037WO f-ii), or a salt, stereoisomer,
Figure imgf000094_0001
some sequencing comprising the LNA moiety having a structure of Formula (II-f) is selected from: or
Figure imgf000094_0002
In some embodiments, the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-g): a salt, stereoisomer, tautomer, or hydrate thereof,
Figure imgf000094_0003
B is a nucleobase; each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 Attorney Docket No.: F2099-7037WO alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C -C alkyl, C -C B 1 6 1 6-heteroalkyl, cycloalkyl, or heterocyclyl; or R and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; n is 1, 2, 3, 4, 5, or 6; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-g-i) or Formula (II-g-ii): g-ii), or a salt, stereoisomer, tautomer, or
Figure imgf000095_0001
In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-g-iii): iii), or a salt, stereoisomer, tautomer, or hydrate thereof.
Figure imgf000095_0002
contains a nucleotide comprising the LNA moiety has a structure of Formula (II-l): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, O, or N(RB); each of R1, R2, and R3 is independently
Figure imgf000096_0001
C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; M is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the of the TREM. In some embodiments, the TREM contains a nucleotide comprising the LNA moiety having the structure of Formula (II-m): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6
Figure imgf000097_0001
C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R6a and R6b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1- C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety having a structure of Formula (II-l) is selected from: Attorney Docket No.: F2099-7037WO
Figure imgf000098_0001
moiety has a structure of Formula (II-m-i): a salt, stereoisomer, tautomer, or hydrate thereof.
Figure imgf000098_0002
contains a nucleotide comprising the LNA moiety having the structure of has a structure of Formula (II-r): a salt, stereoisomer, tautomer, or hydrate thereof,
Figure imgf000098_0003
or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted Attorney Docket No.: F2099-7037WO with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R6a and R6b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R7a and R7b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1- C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R5a and R5b is independently hydrogen. In an embodiment, each of R5a and R5b is independently C1-C6 alkyl. In an embodiment, each of R5a and R5b is independently C1-C6 heteroalkyl. In an embodiment, each of R5a and R5b is independently C1-C6 haloalkyl. In an embodiment, each of R5a and R5b is independently halo. In an embodiment, one of R5a and R5b is independently hydrogen. In an embodiment, one of R5a and R5b is independently C1-C6 alkyl. In an embodiment, one of R5a and R5b is independently C1-C6 heteroalkyl. In an embodiment, one of R5a and R5b is independently C1-C6 haloalkyl. In an embodiment, one of R5a and R5b is independently halo. Attorney Docket No.: F2099-7037WO In some embodiments, each of R6a and R6b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R6a and R6b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl. In an embodiment, each of R6a and R6b is independently hydrogen. In an embodiment, each of R6a and R6b is independently C1-C6 alkyl. In an embodiment, each of R6a and R6b is independently C1-C6 heteroalkyl. In an embodiment, each of R6a and R6b is independently C1-C6 haloalkyl. In an embodiment, each of R6a and R6b is independently halo. In an embodiment, one of R6a and R6b is independently hydrogen. In an embodiment, one of R6a and R6b is independently C1-C6 alkyl. In an embodiment, one of R6a and R6b is independently C1-C6 heteroalkyl. In an embodiment, one of R6a and R6b is independently C1-C6 haloalkyl. In an embodiment, one of R6a and R6b is independently halo. In some embodiments, the nucleotide within the TREM sequence comprising the LNA moiety has the structure of Formula (II-s) or Formula (II-t): a salt, stereoisomer, tautomer,
Figure imgf000100_0001
In some embodiments, the TREM comprises a single LNA moiety. In some embodiments, the TREM comprises a plurality of LNA moieties. In some embodiments, the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 LNA moieties. In some embodiments, the TREM comprises 2 LNA moieties. In some embodiments, the TREM comprises 3 LNA moieties. In some embodiments, the TREM comprises 4 LNA moieties. In some embodiments, the TREM comprises 5 LNA moieties. In some embodiments, the TREM comprises 6 LNA moieties. In some embodiments, the TREM comprises at least 7 LNA moieties. In some embodiments, the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. In some embodiments, the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. In some embodiments, the LNA moiety is present in the [L2]-[DH Domain]-[L3]. In some embodiments, the LNA moiety is present in the [L3]-[ACH Domain]. In some embodiments, the LNA moiety Attorney Docket No.: F2099-7037WO is present in the [VL Domain]. In some embodiments, the LNA moiety is present in the [TH Domain]. In some embodiments, the LNA moiety is present in the [L4]-[ASt Domain2]-[L5]. In some embodiments, the TREM comprises a locked nucleic acid in the [ASt Domain1]. In some embodiments, the TREM comprises a locked nucleic acid in the [DH Domain]. In some embodiments, the TREM comprises a locked nucleic acid in the [ACH Domain]. In some embodiments, the TREM comprises a locked nucleic acid in the [VL Domain]. In some embodiments, the TREM comprises a locked nucleic acid in the [TH Domain]. In some embodiments, the TREM comprises a locked nucleic acid in the [ASt Domain2]. In an embodiment, the LNA moiety is present at any one of nucleotide positions 1, 14, 15, 16, 28, 29, 43, 44, 45, 54, 65, 68, and 70 of the TREM. In an embodiment, the locked nucleic acid is present at any one of nucleotide position 14, 15, 44, or 54 of the TREM. In an embodiment, the locked nucleic acid is not present at one of the nucleotide positions 41, 42, or 63 of the TREM. In an embodiment, the locked nucleic acid is not present at the nucleotide position 26 of the TREM. In some embodiments, the TREM comprises two or more locked nucleic acids. In some embodiments, the TREM comprises two locked nucleic acids. In some embodiments, the TREM comprises two locked nucleic acids, each independently located in the [ASt Domain1] and the [VL Domain]. In some embodiments, the TREM comprises two locked nucleic acids, each independently located in the [ASt Domain1] and the [TH Domain]. In some embodiments, the TREM comprises two locked nucleic acids, each independently located in the [ASt Domain1] and the [ASt Domain2]. In some embodiments, the TREM comprises two locked nucleic acids, each independently located at one of nucleotide positions 14, 15, 44, 54, and 65 (e.g., 14 and 15, e.g., 15 and 65, e.g., 14 and 65, or e.g., 14 and 54). In some embodiments, the TREM comprises three locked nucleic acids. In some embodiments, the TREM comprises three locked nucleic acids each independently located in the [ACH Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM comprises one locked nucleic acid located in the [ACH Domain], and two locked nucleic acids in the [VL Domain]. In some embodiments, the TREM comprises one locked nucleic acid in the [TH Domain], and two locked nucleic acids in the [DH Domain]. In some embodiments, the TREM comprises three locked nucleic acids, each independently located at any of the nucleotide positions 14, 15, 54, or 65 (e.g., 14, 15, and 54, e.g., 14, 15, and 65). In some embodiments, the Attorney Docket No.: F2099-7037WO TREM comprises three locked nucleic acids, each independently located at nucleotide position 14, 15, and 54. In some embodiments, the TREM comprises three locked nucleic acids, each independently located at nucleotide position 14, 15, and 65. In some embodiments, the TREM comprises four locked nucleic acids located in the [DH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM comprises four locked nucleic acids located in the [DH Domain], the [ACH Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM comprises four locked nucleic acids, each independently located at nucleotide position 14, 44, 54, and 65. In some embodiments, the TREM comprises four locked nucleic acids, each independently located at nucleotide position 14, 43, 54, and 65. In some embodiments, the TREM comprises five locked nucleic acids. In some embodiments, the TREM comprises six locked nucleic acids. In some embodiments, the TREM comprises seven or more locked nucleic acids. In some embodiments, the TREM is a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 625-793, 992-1252, 1357, 1364, 1378-1393, or 1397-1398 (e.g., any one of SEQ ID Nos: 625-793, e.g., any one of SEQ ID Nos: 992-1252, 1357, 1364, 1378-1393, or 1397-1398). In some embodiment, the TREM is a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793, 992-1252, 1357, 1364, 1378- 1393, and 1397-1398 (e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793, or a TREM having a sequence of any one of SEQ ID NOs: 992-1252, 1357, 1364, 1378-1393, and 1397-1398). In some embodiment, the TREM is a TREM having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or more sequence identity to a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793. In some embodiments, the TREM is a TREM having at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 1617, 18, 19, 20, or more nucleotides that differ from a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793. In some embodiments, the TREM is a TREM having 1-5, 5-10, 10-15, or 15-20 nucleotides that differ from a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793. In some embodiments, the TREM is a TREM having at least one additional non-naturally occurring modification compared with a TREM provided in FIG.1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793. In some embodiments, the TREM is a TREM having at least 2, 3, 4, 5, 6, 7, 8.9, 10, or Attorney Docket No.: F2099-7037WO more additional non-naturally occurring modifications compared with a TREM provided in FIG. 1, e.g., a TREM having a sequence of any one of SEQ ID NOs: 626-793. In an embodiment, the locked nucleic acid is located at the nucleotide position 14, 15, 44, 54, or 65 of the TREM. In an embodiment, the locked nucleic acid is located at the nucleotide position 14 of the TREM. In an embodiment, the locked nucleic acid is located at nucleotide position 15 of the TREM. In an embodiment, the locked nucleic acid is located at the nucleotide position 54 of the TREM. In an embodiment, the locked nucleic acid is located at the nucleotide position 65 of the TREM. In an embodiment, the locked nucleic acid is not located at the nucleotide position 41, 42, or 63 of the TREM. In an embodiment, the locked nucleic acid is not located at the nucleotide position 7, 8, 9, 10, 17, 18, 31, 32, 33, 34, 35, 36, 37, 38, 39, 48, 55, 58, 59, 60, 61, 72, or 73 (e.g., 7-10, 17-18, 31-39, 48, 55, 58-61, or 72-73). In an embodiment, the locked nucleic acid is not located at the nucleotide positions 7-10 (e.g., 7, 8, 9, or 10). In an embodiment, the locked nucleic acid is not located at the nucleotide position 7. In an embodiment, the locked nucleic acid is not located at the nucleotide position 8. In an embodiment, the locked nucleic acid is not located at the nucleotide position 9. In an embodiment, the locked nucleic acid is not located at the nucleotide position 10. In an embodiment, the locked nucleic acid is not located at the nucleotide position 17-18 (e.g., 17 or 18). In an embodiment, the locked nucleic acid is not located at the nucleotide position 17. In an embodiment, the locked nucleic acid is not located at the nucleotide position 18. In an embodiment, the locked nucleic acid is not located at the nucleotide position 31-39 (e.g., 31, 32, 33, 34, 35, 36, 37, 38, or 39). In an embodiment, the locked nucleic acid is not located at the nucleotide position 31. In an embodiment, the locked nucleic acid is not located at the nucleotide position 32. In an embodiment, the locked nucleic acid is not located at the nucleotide position 33. In an embodiment, the locked nucleic acid is not located at the nucleotide position 34. In an embodiment, the locked nucleic acid is not located at the nucleotide position 35. In an embodiment, the locked nucleic acid is not located at the nucleotide position 36. In an embodiment, the locked nucleic acid is not located at the nucleotide position 37. In an embodiment, the locked nucleic acid is not located at the nucleotide position 38. In an embodiment, the locked nucleic acid is not located at the nucleotide position 39. Attorney Docket No.: F2099-7037WO In an embodiment, the locked nucleic acid is not located at the nucleotide position 48. In an embodiment, the locked nucleic acid is not located at the nucleotide position 55. In an embodiment, the locked nucleic acid is not located at the nucleotide position 58-61 (e.g., 58, 59, 60, or 61). In an embodiment, the locked nucleic acid is not located at the nucleotide position 58. In an embodiment, the locked nucleic acid is not located at the nucleotide position 59. In an embodiment, the locked nucleic acid is not located at the nucleotide position 60. In an embodiment, the locked nucleic acid is not located at the nucleotide position 61. In an embodiment, the locked nucleic acid is not located at the nucleotide position 72-73 (e.g., 72 or 73). In an embodiment, the locked nucleic acid is not located at the nucleotide position 72. In an embodiment, the locked nucleic acid is not located at the nucleotide position 73. In some embodiments, the TREM comprises the non-naturally occurring modification pattern of TREM No.166 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 12-m, 13-f, 14-LNA, 16-m, 17-m, 19-*, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 41-m, 42- m, 43-LNA, 44-f, 45-LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 60- f, 61-m, 62-m, 63-f*, 64-d, 65-LNA, 66-m, 67-m, 68-f, 71-m, 72-f, 73-m, 75-f*. In some embodiments, the TREM comprises the non-naturally occurring modification pattern of TREM No.167 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 12-m, 13-f, 14-LNA, 15-f, 16-m, 17-m, 19-*, 21-f, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 40-f, 41-m, 42-m, 43-LNA, 44-f, 45-LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 59-f, 60-f, 61-m, 62-m, 63-f*, 64-d, 65-LNA, 66-m, 67-m, 68-f, 71-m, 72-f, 73-m, 75-f*. In some embodiments, the TREM comprises the non-naturally occurring modification pattern of TREM No.168 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 13-f, 15-f, 17- m, 19-*, 20-d, 21-f, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 40-f, 41-m, 42-m, 43-LNA, 44-f, 45-LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 59-f, 60-f, 61-m, 62-m, 63-f*, 64-d, 65-LNA, 66-m, 67-m, 68-f, 72-f, 73-m, 75-f*. In some embodiments, the TREM comprises the non-naturally occurring modification pattern of TREM No.169 of FIG.1: 1-m*, 2-m, 3-m, 4-m, 5-m, 6-m, 10-m, 11-m, 13-f, 17-m, 19-*, 22-m, 25-m, 27-m*, 28-f, 29-moe, 33-f, 35-*, 37-*, 38-*, 41-m, 42-m, 43-LNA, 44-f, 45- Attorney Docket No.: F2099-7037WO LNA, 46-d, 49-m, 50-m, 51-m, 52-d, 53-m, 54-LNA, 55-*, 56-m, 57-f, 60-f, 61-m, 62-m, 63-f*, 64-d, 65-LNA, 66-m, 67-m, 68-f, 72-f, 73-m, 75-f*. In some embodiments, the TREM containing an LNA moiety is TREM No.15. In some embodiments, the TREM containing an LNA moiety is TREM No.16. In some embodiments, the TREM containing an LNA moiety is TREM No.43. In some embodiments, the TREM containing an LNA moiety is TREM No.44. In some embodiments, the TREM containing an LNA moiety is TREM No.45. In some embodiments, the TREM containing an LNA moiety is TREM No.46. In some embodiments, the TREM containing an LNA moiety is TREM No.66. In some embodiments, the TREM containing an LNA moiety is TREM No.79. In some embodiments, the TREM containing an LNA moiety is TREM No.80. In some embodiments, the TREM containing an LNA moiety is TREM No.81. In some embodiments, the TREM containing an LNA moiety is TREM No.82. In some embodiments, the TREM containing an LNA moiety is TREM No.83. In some embodiments, the TREM containing an LNA moiety is TREM No.84. In some embodiments, the TREM containing an LNA moiety is TREM No.85. In some embodiments, the TREM containing an LNA moiety is TREM No.86. In some embodiments, the TREM containing an LNA moiety is TREM No.87. In some embodiments, the TREM containing an LNA moiety is TREM No.88. In some embodiments, the TREM containing an LNA moiety is TREM No.89. In some embodiments, the TREM containing an LNA moiety is TREM No.90. In some embodiments, the TREM containing an LNA moiety is TREM No.91. In some embodiments, the TREM containing an LNA moiety is TREM No.92. In some embodiments, the TREM containing an LNA moiety is TREM No.93. In some embodiments, the TREM containing an LNA moiety is TREM No.94. In some embodiments, the TREM containing an LNA moiety is TREM No.95. In some embodiments, the TREM containing an LNA moiety is TREM No.96. In some embodiments, the TREM containing an LNA moiety is TREM No.97. In some embodiments, the TREM containing an LNA moiety is TREM No.98. In some embodiments, the TREM containing an LNA moiety is TREM No.99. In some embodiments, the TREM containing an LNA moiety is TREM No.100. In some embodiments, the TREM containing an LNA moiety is TREM No.101. In some embodiments, the TREM containing an LNA moiety is TREM No.102. In some embodiments, the TREM containing an LNA moiety is TREM No.103. In some embodiments, the TREM containing an LNA moiety is TREM No.104. In some embodiments, the TREM containing an LNA moiety is Attorney Docket No.: F2099-7037WO TREM No.105. In some embodiments, the TREM containing an LNA moiety is TREM No.106. In some embodiments, the TREM containing an LNA moiety is TREM No.107. In some embodiments, the TREM containing an LNA moiety is TREM No.108. In some embodiments, the TREM containing an LNA moiety is TREM No.109. In some embodiments, the TREM containing an LNA moiety is TREM No.110. In some embodiments, the TREM containing an LNA moiety is TREM No.111. In some embodiments, the TREM containing an LNA moiety is TREM No.112. In some embodiments, the TREM containing an LNA moiety is TREM No.113. In some embodiments, the TREM containing an LNA moiety is TREM No.114. In some embodiments, the TREM containing an LNA moiety is TREM No.115. In some embodiments, the TREM containing an LNA moiety is TREM No.116. In some embodiments, the TREM containing an LNA moiety is TREM No.117. In some embodiments, the TREM containing an LNA moiety is TREM No.118. In some embodiments, the TREM containing an LNA moiety is TREM No.120. In some embodiments, the TREM containing an LNA moiety is TREM No.121. In some embodiments, the TREM containing an LNA moiety is TREM No.122. In some embodiments, the TREM containing an LNA moiety is TREM No.123. In some embodiments, the TREM containing an LNA moiety is TREM No.124. In some embodiments, the TREM containing an LNA moiety is TREM No.125. In some embodiments, the TREM containing an LNA moiety is TREM No.126. In some embodiments, the TREM containing an LNA moiety is TREM No.127. In some embodiments, the TREM containing an LNA moiety is TREM No.128. In some embodiments, the TREM containing an LNA moiety is TREM No.129. In some embodiments, the TREM containing an LNA moiety is TREM No.130. In some embodiments, the TREM containing an LNA moiety is TREM No.132. In some embodiments, the TREM containing an LNA moiety is TREM No.133. In some embodiments, the TREM containing an LNA moiety is TREM No.134. In some embodiments, the TREM containing an LNA moiety is TREM No.135. In some embodiments, the TREM containing an LNA moiety is TREM No.136. In some embodiments, the TREM containing an LNA moiety is TREM No.137. In some embodiments, the TREM containing an LNA moiety is TREM No.138. In some embodiments, the TREM containing an LNA moiety is TREM No.139. In some embodiments, the TREM containing an LNA moiety is TREM No.140. In some embodiments, the TREM containing an LNA moiety is TREM No.141. In some embodiments, the TREM containing an LNA moiety is TREM No.142. In some embodiments, the TREM containing an LNA moiety is TREM No.143. Attorney Docket No.: F2099-7037WO In some embodiments, the TREM containing an LNA moiety is TREM No.144. In some embodiments, the TREM containing an LNA moiety is TREM No.145. In some embodiments, the TREM containing an LNA moiety is TREM No.146. In some embodiments, the TREM containing an LNA moiety is TREM No.147. In some embodiments, the TREM containing an LNA moiety is TREM No.148. In some embodiments, the TREM containing an LNA moiety is TREM No.149. In some embodiments, the TREM containing an LNA moiety is TREM No.150. In some embodiments, the TREM containing an LNA moiety is TREM No.151. In some embodiments, the TREM containing an LNA moiety is TREM No.152. In some embodiments, the TREM containing an LNA moiety is TREM No.153. In some embodiments, the TREM containing an LNA moiety is TREM No.154. In some embodiments, the TREM containing an LNA moiety is TREM No.155. In some embodiments, the TREM containing an LNA moiety is TREM No.156. In some embodiments, the TREM containing an LNA moiety is TREM No.157. In some embodiments, the TREM containing an LNA moiety is TREM No.158. In some embodiments, the TREM containing an LNA moiety is TREM No.159. In some embodiments, the TREM containing an LNA moiety is TREM No.160. In some embodiments, the TREM containing an LNA moiety is TREM No.161. In some embodiments, the TREM containing an LNA moiety is TREM No.162. In some embodiments, the TREM containing an LNA moiety is TREM No.163. In some embodiments, the TREM containing an LNA moiety is TREM No.164. In some embodiments, the TREM containing an LNA moiety is TREM No.165. In some embodiments, the TREM containing an LNA moiety is TREM No.166. In some embodiments, the TREM containing an LNA moiety is TREM No.444. In some embodiments, the TREM containing an LNA moiety is TREM No.445. In some embodiments, the TREM containing an LNA moiety is TREM No.451. In some embodiments, the TREM containing an LNA moiety is TREM No.454. In some embodiments, the TREM containing an LNA moiety is TREM No.547. In some embodiments, the TREM containing an LNA moiety is TREM No.566. In some embodiments, the TREM containing an LNA moiety is TREM No.628. 2’,5’-Linked Nucleotides In one aspect, the present disclosure consists of a TREM sequence comprising the 2’,5’- linked nucleotide having a structure of Formula (III): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, wherein: B is a ; each of R1, R2, and R4 is independently hydrogen, C1-C6
Figure imgf000108_0001
C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or - N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2- C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7- membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom,
Figure imgf000108_0002
of the TREM. In some embodiments, B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). In some embodiments, B is a purine-based nucleobase. In some embodiments, B is a pyrimidine-based nucleobase. In some embodiments, B is adenine. In some embodiments, B is cytosine. In some embodiments, B is guanine. In some embodiments, B is uracil. In some embodiments, B is a non-naturally occurring nucleobase. In some embodiments, B comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. In some embodiments, the chemical Attorney Docket No.: F2099-7037WO modification is a nucleotide sugar modification. In some embodiments, the chemical modification is a 2’OMe modification. In some embodiments, the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification. In some embodiments, the chemical modification is a 2’MOE modification. In some embodiments, the chemical modification is a 2’-deoxy modification. In some embodiments, the chemical modification is present in the internucleotide region (e.g., a backbone modification). In some embodiments, the chemical modification is a phosphorothioate modification. In some embodiments, B does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(RB). In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R1 is hydrogen. In some embodiments, R1 is C1-C6 alkyl. In some embodiments, R1 is C2-C6 alkenyl. In some embodiments, R1 is C2-C6 alkynyl. In some embodiments, R1 is C1- C6 heteroalkyl. In some embodiments, R1 is C1-C6 haloalkyl. In some embodiments, R1 is halo. In some embodiments, R1 is cyano. In some embodiments, R1 is -ORA. In some embodiments, R1 is -N(RB)(RC). In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R2 is hydrogen. In some embodiments, R2 is C1-C6 alkyl. In some embodiments, R2 is C2-C6 alkenyl. In some embodiments, R2 is C2-C6 alkynyl. In some embodiments, R2 is C1- C6 heteroalkyl. In some embodiments, R2 is C1-C6 haloalkyl. In some embodiments, R2 is halo. In some embodiments, R2 is cyano. In some embodiments, R2 is -ORA. In some embodiments, R2 is -N(RB)(RC). In some embodiments, R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R4 is hydrogen. In some embodiments, R4 is C1-C6 alkyl. In some embodiments, R4 is C2-C6 alkenyl. In some embodiments, R4 is C2-C6 alkynyl. In some embodiments, R4 is C1- C 6 heteroalkyl. In some embodiments, R4 is C 1 -C 6 haloalkyl. In some embodiments, R4 is halo. Attorney Docket No.: F2099-7037WO In some embodiments, R4 is cyano. In some embodiments, R4 is -ORA. In some embodiments, R4 is -N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently hydrogen. In some embodiments, one of R3a and R3b is independently C1-C6 alkyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, one of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, one of R3a and R3b is independently halo. In some embodiments, one of R3a and R3b is independently cyano. In some embodiments, one of R3a and R3b is independently -ORA. In some embodiments, one of R3a and R3b is independently - N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen. In some embodiments, each of R3a and R3b is independently C1-C6 alkyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, each of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, each of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, each of R3a and R3b is independently halo. In some embodiments, each of R3a and R3b is independently cyano. In some embodiments, each of R3a and R3b is independently -ORA. In some embodiments, each of R3a and R3b is independently -N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently hydrogen. In some embodiments, one of R5a and R5b is independently C1-C6 alkyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, one of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, one of R5a and R5b is independently halo. In some embodiments, one of R5a and R5b is independently cyano. In some embodiments, one of Attorney Docket No.: F2099-7037WO R5a and R5b is independently -ORA. In some embodiments, one of R5a and R5b is independently - N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen. In some embodiments, each of R5a and R5b is independently C1-C6 alkyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, each of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, each of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, each of R5a and R5b is independently halo. In some embodiments, each of R5a and R5b is independently cyano. In some embodiments, each of R5a and R5b is independently -ORA. In some embodiments, each of R5a and R5b is independently -N(RB)(RC). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C1-C6 alkyl. In some embodiments, each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen, C1- C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen or C1- C6 alkyl. In some embodiments, each RA is independently hydrogen. In some embodiments, each of RB and RC is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of RB and RC is independently hydrogen or C1-C6 alkyl. In some embodiments, each of RB and RC is independently hydrogen. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 alkyl. In some embodiments, R10 is C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 haloalkyl. In some embodiments, R10 is halo. In some embodiments, R10 is cyano. In some embodiments, R10 is -ORA1. In some embodiments, RA1 is hydrogen. In some embodiments, RA1 is C1-C6 alkyl. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, TREM comprises a single 2’,5’-linked nucleotide. In some embodiments, the TREM comprises a plurality of 2’,5’-linked nucleotides. In some Attorney Docket No.: F2099-7037WO embodiments, the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 252’,5’-linked nucleotides. In some embodiments, the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]- [L2]. In some embodiments, the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]- [L2]. In some embodiments, the 2’,5’-linked nucleotide is present in the [L2]-[DH Domain]- [L3]. In some embodiments, the 2’,5’-linked nucleotide is present in the [L3]-[ACH Domain]. In some embodiments, the 2’,5’-linked nucleotide is present in the [VL Domain]. In some embodiments, the 2’,5’-linked nucleotide is present in the [TH Domain]. In some embodiments, the 2’,5’-linked nucleotide is present in the [L4]-[ASt Domain2]-[L5]. In some embodiments, the TREM has a sequence selected from a sequence provided in FIG.1. In some embodiments, the TREM comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in Table 1. In some embodiments, the TREM comprises a nucleotide sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from the nucleotide sequence of a TREM provided in Table 1, e.g., any one of the TREMs having SEQ ID NO.794-991, 1060, 1062, 1112, 1135, 1153-1158, 1161, 1163, 1166-1170, 1175, 1200, 1207, and 1253-1415 (e.g., any one of the TREMs having SEQ ID NO.794-991, e.g., any one of the TREMs having SEQ ID NO. 1060, 1062, 1112, 1135, 1153-1158, 1161, 1163, 1166-1170, 1175, 1200, 1207, and 1253-1415). In some embodiments, the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non- naturally occurring modifications compared with a TREM provided in Table 1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications). In some embodiments, the TREM comprises the nucleotide sequence of any one of the TREMs having SEQ ID NO.794-991, 1060, 1062, 1112, 1135, 1153- 1158, 1161, 1163, 1166-1170, 1175, 1200, 1207, or 1253-1415 (e.g., any one of the TREMs having SEQ ID NO.794-991. In one aspect, the present disclosure consists of a TREM sequence comprising the 2’,5’- linked nucleotide having a structure of Formula (III-a): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, or N(RB); each of R1, R2, and R4 is independently
Figure imgf000113_0001
C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R3a is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R3c is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2- C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2- C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. In some embodiments, B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). In some embodiments, B is a non-naturally occurring nucleobase. In some embodiments, B comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. In some embodiments, the chemical modification is a nucleotide sugar modification. In some embodiments, the chemical Attorney Docket No.: F2099-7037WO modification is a 2’OMe modification. In some embodiments, the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification. In some embodiments, the chemical modification is a 2’MOE modification. In some embodiments, the chemical modification is a 2’-deoxy modification. In some embodiments, the chemical modification is present in the internucleotide region (e.g., a backbone modification). In some embodiments, the chemical modification is a phosphorothioate modification. In some embodiments, B does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). In some embodiments, Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(RB). In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R1 is hydrogen. In some embodiments, R1 is C1-C6 alkyl. In some embodiments, R1 is C2-C6 alkenyl. In some embodiments, R1 is C2-C6 alkynyl. In some embodiments, R1 is C1- C6 heteroalkyl. In some embodiments, R1 is C1-C6 haloalkyl. In some embodiments, R1 is halo. In some embodiments, R1 is cyano. In some embodiments, R1 is -ORA. In some embodiments, R1 is -N(RB)(RC). In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R2 is hydrogen. In some embodiments, R2 is C1-C6 alkyl. In some embodiments, R2 is C2-C6 alkenyl. In some embodiments, R2 is C2-C6 alkynyl. In some embodiments, R2 is C1- C6 heteroalkyl. In some embodiments, R2 is C1-C6 haloalkyl. In some embodiments, R2 is halo. In some embodiments, R2 is cyano. In some embodiments, R2 is -ORA. In some embodiments, R2 is -N(RB)(RC). In some embodiments, R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R4 is hydrogen. In some embodiments, R4 is C1-C6 alkyl. In some embodiments, R4 is C2-C6 alkenyl. In some embodiments, R4 is C2-C6 alkynyl. In some embodiments, R4 is C1- C6 heteroalkyl. In some embodiments, R4 is C1-C6 haloalkyl. In some embodiments, R4 is halo. In some embodiments, R4 is cyano. In some embodiments, R4 is -ORA. In some embodiments, R4 is -N(RB)(RC). Attorney Docket No.: F2099-7037WO In some embodiments, R3a is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1- C6 haloalkyl, or halo. In some embodiments, R3a is independently hydrogen, C1-C6 alkyl, or C1- C6 heteroalkyl. In some embodiments, R3a is hydrogen. In some embodiments, R3a is C1-C6 alkyl. In some embodiments, R3a is C2-C6 alkenyl. In some embodiments, R3a is C2-C6 alkynyl. In some embodiments, R3a is C1-C6 heteroalkyl. In some embodiments, R3a is C1-C6 haloalkyl. In some embodiments, R3a is halo. In some embodiments, R3a is cyano. In some embodiments, R3a is - ORA. In some embodiments, R3a is -N(RB)(RC). In some embodiments, R3c is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10. In some embodiments, R3c is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R3c is hydrogen. In some embodiments, R3c is C1-C6 alkyl. In some embodiments, R3c is C1-C6 heteroalkyl. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently hydrogen. In some embodiments, one of R5a and R5b is independently C1-C6 alkyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, one of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, one of R5a and R5b is independently halo. In some embodiments, one of R5a and R5b is independently cyano. In some embodiments, one of R5a and R5b is independently -ORA. In some embodiments, one of R5a and R5b is independently - N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen. In some embodiments, each of R5a and R5b is independently C1-C6 alkyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, each of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, each of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, each of R5a and R5b is independently halo. In some embodiments, each of R5a and R5b is independently cyano. In some embodiments, each of R5a and R5b is independently -ORA. In some embodiments, each of R5a and R5b is independently -N(RB)(RC). Attorney Docket No.: F2099-7037WO In some embodiments, R’ is hydrogen. In some embodiments, R’ is C1-C6 alkyl. In some embodiments, each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen, C1- C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen or C1- C6 alkyl. In some embodiments, each RA is independently hydrogen. In some embodiments, each of RB and RC is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of RB and RC is independently hydrogen or C1-C6 alkyl. In some embodiments, each of RB and RC is independently hydrogen. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 alkyl. In some embodiments, R10 is C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 haloalkyl. In some embodiments, R10 is halo. In some embodiments, R10 is cyano. In some embodiments, R10 is -ORA1. In some embodiments, RA1 is hydrogen. In some embodiments, RA1 is C1-C6 alkyl. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, the nucleotide within the TREM sequence comprising the 2’,5’- linked nucleotide has a structure of Formula (III-a-i) or Formula (III-a-ii):
Figure imgf000116_0001
In one aspect, the present disclosure consists of a TREM sequence comprising the 2’,5’- linked nucleotide having a structure of Formula (III-b): Attorney Docket No.: F2099-7037WO a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y and R4 is independently hydrogen, C1-C6 alkyl, C2-C6
Figure imgf000117_0001
C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C -C alkyl, C -C - B 1 6 1 6 heteroalkyl, cycloalkyl, or heterocyclyl; or R and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom,
Figure imgf000117_0002
of the TREM. In some embodiments, Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(RB). In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R1 is hydrogen. In some embodiments, R1 is C1-C6 alkyl. In some embodiments, Attorney Docket No.: F2099-7037WO R1 is C2-C6 alkenyl. In some embodiments, R1 is C2-C6 alkynyl. In some embodiments, R1 is C1- C6 heteroalkyl. In some embodiments, R1 is C1-C6 haloalkyl. In some embodiments, R1 is halo. In some embodiments, R1 is cyano. In some embodiments, R1 is -ORA. In some embodiments, R1 is -N(RB)(RC). In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R2 is hydrogen. In some embodiments, R2 is C1-C6 alkyl. In some embodiments, R2 is C2-C6 alkenyl. In some embodiments, R2 is C2-C6 alkynyl. In some embodiments, R2 is C1- C6 heteroalkyl. In some embodiments, R2 is C1-C6 haloalkyl. In some embodiments, R2 is halo. In some embodiments, R2 is cyano. In some embodiments, R2 is -ORA. In some embodiments, R2 is -N(RB)(RC). In some embodiments, R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R4 is hydrogen. In some embodiments, R4 is C1-C6 alkyl. In some embodiments, R4 is C2-C6 alkenyl. In some embodiments, R4 is C2-C6 alkynyl. In some embodiments, R4 is C1- C6 heteroalkyl. In some embodiments, R4 is C1-C6 haloalkyl. In some embodiments, R4 is halo. In some embodiments, R4 is cyano. In some embodiments, R4 is -ORA. In some embodiments, R4 is -N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently hydrogen. In some embodiments, one of R3a and R3b is independently C1-C6 alkyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, one of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, one of R3a and R3b is independently halo. In some embodiments, one of R3a and R3b is independently cyano. In some embodiments, one of R3a and R3b is independently -ORA. In some embodiments, one of R3a and R3b is independently - N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen. In some embodiments, each of R3a and R3b is independently C1-C6 alkyl. In some embodiments, each of Attorney Docket No.: F2099-7037WO R3a and R3b is independently C2-C6 alkenyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, each of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, each of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, each of R3a and R3b is independently halo. In some embodiments, each of R3a and R3b is independently cyano. In some embodiments, each of R3a and R3b is independently -ORA. In some embodiments, each of R3a and R3b is independently -N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently hydrogen. In some embodiments, one of R5a and R5b is independently C1-C6 alkyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, one of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, one of R5a and R5b is independently halo. In some embodiments, one of R5a and R5b is independently cyano. In some embodiments, one of R5a and R5b is independently -ORA. In some embodiments, one of R5a and R5b is independently - N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen. In some embodiments, each of R5a and R5b is independently C1-C6 alkyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, each of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, each of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, each of R5a and R5b is independently halo. In some embodiments, each of R5a and R5b is independently cyano. In some embodiments, each of R5a and R5b is independently -ORA. In some embodiments, each of R5a and R5b is independently -N(RB)(RC). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C1-C6 alkyl. In some embodiments, each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen, C1- C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen or C1- C6 alkyl. In some embodiments, each RA is independently hydrogen. Attorney Docket No.: F2099-7037WO In some embodiments, each of RB and RC is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of RB and RC is independently hydrogen or C1-C6 alkyl. In some embodiments, each of RB and RC is independently hydrogen. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 alkyl. In some embodiments, R10 is C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 haloalkyl. In some embodiments, R10 is halo. In some embodiments, R10 is cyano. In some embodiments, R10 is -ORA1. In some embodiments, RA1 is hydrogen. In some embodiments, RA1 is C1-C6 alkyl. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of Ria and Rib is independently hydrogen or C1-C6 alkyl. In some embodiments, each of Ria and Rib is independently hydrogen. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, or C1- C6 heteroalkyl. In some embodiments, each of Rii and Riii is independently hydrogen or C1-C6 alkyl. In some embodiments, each of Rii and Riii is independently hydrogen. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2. In another aspect, the present disclosure consists of a TREM sequence comprising the 2’,5’-linked nucleotide having a structure of Formula (III-c): a salt, stereoisomer, tautomer, or hydrate
Figure imgf000120_0001
R2, and R4 is independently hydrogen, C1- C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally Attorney Docket No.: F2099-7037WO substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2- C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7- membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, or C1-C6 haloalkyl; Rii is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl; Riii is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. In some embodiments, Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(RB). In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R1 is hydrogen. In some embodiments, R1 is C1-C6 alkyl. In some embodiments, R1 is C2-C6 alkenyl. In some embodiments, R1 is C2-C6 alkynyl. In some embodiments, R1 is C1- C6 heteroalkyl. In some embodiments, R1 is C1-C6 haloalkyl. In some embodiments, R1 is halo. In some embodiments, R1 is cyano. In some embodiments, R1 is -ORA. In some embodiments, R1 is -N(RB)(RC). In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R2 is hydrogen. In some embodiments, R2 is C1-C6 alkyl. In some embodiments, R2 is C2-C6 alkenyl. In some embodiments, R2 is C2-C6 alkynyl. In some embodiments, R2 is C1- C6 heteroalkyl. In some embodiments, R2 is C1-C6 haloalkyl. In some embodiments, R2 is halo. Attorney Docket No.: F2099-7037WO In some embodiments, R2 is cyano. In some embodiments, R2 is -ORA. In some embodiments, R2 is -N(RB)(RC). In some embodiments, R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R4 is hydrogen. In some embodiments, R4 is C1-C6 alkyl. In some embodiments, R4 is C2-C6 alkenyl. In some embodiments, R4 is C2-C6 alkynyl. In some embodiments, R4 is C1- C6 heteroalkyl. In some embodiments, R4 is C1-C6 haloalkyl. In some embodiments, R4 is halo. In some embodiments, R4 is cyano. In some embodiments, R4 is -ORA. In some embodiments, R4 is -N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently hydrogen. In some embodiments, one of R3a and R3b is independently C1-C6 alkyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, one of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, one of R3a and R3b is independently halo. In some embodiments, one of R3a and R3b is independently cyano. In some embodiments, one of R3a and R3b is independently -ORA. In some embodiments, one of R3a and R3b is independently - N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen. In some embodiments, each of R3a and R3b is independently C1-C6 alkyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, each of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, each of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, each of R3a and R3b is independently halo. In some embodiments, each of R3a and R3b is independently cyano. In some embodiments, each of R3a and R3b is independently -ORA. In some embodiments, each of R3a and R3b is independently -N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C 1 -C 6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R5a Attorney Docket No.: F2099-7037WO and R5b is independently hydrogen. In some embodiments, one of R5a and R5b is independently C1-C6 alkyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, one of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, one of R5a and R5b is independently halo. In some embodiments, one of R5a and R5b is independently cyano. In some embodiments, one of R5a and R5b is independently -ORA. In some embodiments, one of R5a and R5b is independently - N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen. In some embodiments, each of R5a and R5b is independently C1-C6 alkyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, each of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, each of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, each of R5a and R5b is independently halo. In some embodiments, each of R5a and R5b is independently cyano. In some embodiments, each of R5a and R5b is independently -ORA. In some embodiments, each of R5a and R5b is independently -N(RB)(RC). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C1-C6 alkyl. In some embodiments, each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen, C1- C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen or C1- C6 alkyl. In some embodiments, each RA is independently hydrogen. In some embodiments, each of RB and RC is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of RB and RC is independently hydrogen or C1-C6 alkyl. In some embodiments, each of RB and RC is independently hydrogen. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 alkyl. In some embodiments, R10 is C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 haloalkyl. In some embodiments, R10 is halo. In some embodiments, R10 is cyano. In some embodiments, R10 is -ORA1. In some embodiments, RA1 is hydrogen. In some embodiments, RA1 is C1-C6 alkyl. Attorney Docket No.: F2099-7037WO In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of Ria and Rib is independently hydrogen or C1-C6 alkyl. In some embodiments, each of Ria and Rib is independently hydrogen. In some embodiments, Rii is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, Rii is hydrogen or C1-C6 alkyl. In some embodiments, Rii is hydrogen. In some embodiments, Riii is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, Riii is hydrogen or C1-C6 alkyl. In some embodiments, Riii is hydrogen. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2. In another aspect, the present disclosure consists of a TREM sequence comprising the 2’,5’-linked nucleotide having a structure of Formula (III-d): a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y
Figure imgf000124_0001
and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB Attorney Docket No.: F2099-7037WO and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, of the TREM.
Figure imgf000125_0001
In some embodiments, Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(RB). In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R1 is hydrogen. In some embodiments, R1 is C1-C6 alkyl. In some embodiments, R1 is C2-C6 alkenyl. In some embodiments, R1 is C2-C6 alkynyl. In some embodiments, R1 is C1- C6 heteroalkyl. In some embodiments, R1 is C1-C6 haloalkyl. In some embodiments, R1 is halo. In some embodiments, R1 is cyano. In some embodiments, R1 is -ORA. In some embodiments, R1 is -N(RB)(RC). In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R2 is hydrogen. In some embodiments, R2 is C1-C6 alkyl. In some embodiments, R2 is C2-C6 alkenyl. In some embodiments, R2 is C2-C6 alkynyl. In some embodiments, R2 is C1- C6 heteroalkyl. In some embodiments, R2 is C1-C6 haloalkyl. In some embodiments, R2 is halo. In some embodiments, R2 is cyano. In some embodiments, R2 is -ORA. In some embodiments, R2 is -N(RB)(RC). In some embodiments, R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R4 is hydrogen. In some embodiments, R4 is C1-C6 alkyl. In some embodiments, R4 is C2-C6 alkenyl. In some embodiments, R4 is C2-C6 alkynyl. In some embodiments, R4 is C1- C6 heteroalkyl. In some embodiments, R4 is C1-C6 haloalkyl. In some embodiments, R4 is halo. In some embodiments, R4 is cyano. In some embodiments, R4 is -ORA. In some embodiments, R4 is -N(RB)(RC). Attorney Docket No.: F2099-7037WO In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently hydrogen. In some embodiments, one of R3a and R3b is independently C1-C6 alkyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, one of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, one of R3a and R3b is independently halo. In some embodiments, one of R3a and R3b is independently cyano. In some embodiments, one of R3a and R3b is independently -ORA. In some embodiments, one of R3a and R3b is independently - N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen. In some embodiments, each of R3a and R3b is independently C1-C6 alkyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, each of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, each of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, each of R3a and R3b is independently halo. In some embodiments, each of R3a and R3b is independently cyano. In some embodiments, each of R3a and R3b is independently -ORA. In some embodiments, each of R3a and R3b is independently -N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently hydrogen. In some embodiments, one of R5a and R5b is independently C1-C6 alkyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, one of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, one of R5a and R5b is independently halo. In some embodiments, one of R5a and R5b is independently cyano. In some embodiments, one of R5a and R5b is independently -ORA. In some embodiments, one of R5a and R5b is independently - N(RB)(RC). Attorney Docket No.: F2099-7037WO In some embodiments, each of R5a and R5b is independently hydrogen. In some embodiments, each of R5a and R5b is independently C1-C6 alkyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, each of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, each of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, each of R5a and R5b is independently halo. In some embodiments, each of R5a and R5b is independently cyano. In some embodiments, each of R5a and R5b is independently -ORA. In some embodiments, each of R5a and R5b is independently -N(RB)(RC). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C1-C6 alkyl. In some embodiments, each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen, C1- C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen or C1- C6 alkyl. In some embodiments, each RA is independently hydrogen. In some embodiments, each of RB and RC is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of RB and RC is independently hydrogen or C1-C6 alkyl. In some embodiments, each of RB and RC is independently hydrogen. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 alkyl. In some embodiments, R10 is C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 haloalkyl. In some embodiments, R10 is halo. In some embodiments, R10 is cyano. In some embodiments, R10 is -ORA1. In some embodiments, RA1 is hydrogen. In some embodiments, RA1 is C1-C6 alkyl. In some embodiments, each of Ria and Rib is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of Ria and Rib is independently hydrogen or C1-C6 alkyl. In some embodiments, each of Ria and Rib is independently hydrogen. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, or C1- C6 heteroalkyl. In some embodiments, each of Rii and Riii is independently hydrogen or C1-C6 alkyl. In some embodiments, each of Rii and Riii is independently hydrogen. Attorney Docket No.: F2099-7037WO In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2. In another aspect, the present disclosure consists of a TREM sequence comprising the 2’,5’-linked nucleotide having a structure of Formula (III-e): salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y is and R4 is independently hydrogen, C1-C6 alkyl, C2-C6
Figure imgf000128_0001
alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C -C alkyl, C -C -heteroalkyl B 1 6 1 6 , cycloalkyl, or heterocyclyl; or R and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; Ri is hydrogen, C1-C6 alkyl, C1- C6-heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. Attorney Docket No.: F2099-7037WO In some embodiments, Y is C(R’). In some embodiments, Y is O. In some embodiments, Y is N(RB). In some embodiments, R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R1 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R1 is hydrogen. In some embodiments, R1 is C1-C6 alkyl. In some embodiments, R1 is C2-C6 alkenyl. In some embodiments, R1 is C2-C6 alkynyl. In some embodiments, R1 is C1- C6 heteroalkyl. In some embodiments, R1 is C1-C6 haloalkyl. In some embodiments, R1 is halo. In some embodiments, R1 is cyano. In some embodiments, R1 is -ORA. In some embodiments, R1 is -N(RB)(RC). In some embodiments, R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R2 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R2 is hydrogen. In some embodiments, R2 is C1-C6 alkyl. In some embodiments, R2 is C2-C6 alkenyl. In some embodiments, R2 is C2-C6 alkynyl. In some embodiments, R2 is C1- C6 heteroalkyl. In some embodiments, R2 is C1-C6 haloalkyl. In some embodiments, R2 is halo. In some embodiments, R2 is cyano. In some embodiments, R2 is -ORA. In some embodiments, R2 is -N(RB)(RC). In some embodiments, R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, R4 is hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, R4 is hydrogen. In some embodiments, R4 is C1-C6 alkyl. In some embodiments, R4 is C2-C6 alkenyl. In some embodiments, R4 is C2-C6 alkynyl. In some embodiments, R4 is C1- C6 heteroalkyl. In some embodiments, R4 is C1-C6 haloalkyl. In some embodiments, R4 is halo. In some embodiments, R4 is cyano. In some embodiments, R4 is -ORA. In some embodiments, R4 is -N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R3a and R3b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R3a and R3b is independently hydrogen. In some embodiments, one of R3a and R3b is independently C1-C6 alkyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, one of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, one of R3a and R3b is independently C 1 -C 6 heteroalkyl. In some embodiments, one of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, one of R3a and R3b is independently halo. Attorney Docket No.: F2099-7037WO In some embodiments, one of R3a and R3b is independently cyano. In some embodiments, one of R3a and R3b is independently -ORA. In some embodiments, one of R3a and R3b is independently - N(RB)(RC). In some embodiments, each of R3a and R3b is independently hydrogen. In some embodiments, each of R3a and R3b is independently C1-C6 alkyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkenyl. In some embodiments, each of R3a and R3b is independently C2-C6 alkynyl. In some embodiments, each of R3a and R3b is independently C1-C6 heteroalkyl. In some embodiments, each of R3a and R3b is independently C1-C6 haloalkyl. In some embodiments, each of R3a and R3b is independently halo. In some embodiments, each of R3a and R3b is independently cyano. In some embodiments, each of R3a and R3b is independently -ORA. In some embodiments, each of R3a and R3b is independently -N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. In some embodiments, each of R5a and R5b is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently hydrogen. In some embodiments, one of R5a and R5b is independently C1-C6 alkyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, one of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, one of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, one of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, one of R5a and R5b is independently halo. In some embodiments, one of R5a and R5b is independently cyano. In some embodiments, one of R5a and R5b is independently -ORA. In some embodiments, one of R5a and R5b is independently - N(RB)(RC). In some embodiments, each of R5a and R5b is independently hydrogen. In some embodiments, each of R5a and R5b is independently C1-C6 alkyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkenyl. In some embodiments, each of R5a and R5b is independently C2-C6 alkynyl. In some embodiments, each of R5a and R5b is independently C1-C6 heteroalkyl. In some embodiments, each of R5a and R5b is independently C1-C6 haloalkyl. In some embodiments, each of R5a and R5b is independently halo. In some embodiments, each of R5a and R5b is independently cyano. In some embodiments, each of R5a and R5b is independently -ORA. In some embodiments, each of R5a and R5b is independently -N(RB)(RC). In some embodiments, R’ is hydrogen. In some embodiments, R’ is C1-C6 alkyl. Attorney Docket No.: F2099-7037WO In some embodiments, each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen, C1- C6 alkyl, or C1-C6 heteroalkyl. In some embodiments, each RA is independently hydrogen or C1- C6 alkyl. In some embodiments, each RA is independently hydrogen. In some embodiments, each of RB and RC is independently hydrogen, C1-C6 alkyl or C1- C6-heteroalkyl. In some embodiments, each of RB and RC is independently hydrogen or C1-C6 alkyl. In some embodiments, each of RB and RC is independently hydrogen. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1. In some embodiments, R10 is C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl. In some embodiments, R10 is C1-C6 alkyl or C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 alkyl. In some embodiments, R10 is C1-C6 heteroalkyl. In some embodiments, R10 is C1-C6 haloalkyl. In some embodiments, R10 is halo. In some embodiments, R10 is cyano. In some embodiments, R10 is -ORA1. In some embodiments, RA1 is hydrogen. In some embodiments, RA1 is C1-C6 alkyl. In some embodiments, Ri is hydrogen, C1-C6 alkyl or C1-C6-heteroalkyl. In some embodiments, Ri is hydrogen or C1-C6 alkyl. In some embodiments, Ri is hydrogen. In some embodiments, each of Rii and Riii is independently hydrogen, C1-C6 alkyl, or C1- C6 heteroalkyl. In some embodiments, each of Rii and Riii is independently hydrogen or C1-C6 alkyl. In some embodiments, each of Rii and Riii is independently hydrogen. In some embodiments, each of x and y is independently 0, 1, or 2; In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease. A TREM may comprise multiple 2’,5’-linked nucleotides independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM comprises one 2’,5’-linked nucleotide. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [ASt Domain1]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [DH Domain]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [ACH Domain]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [VL Attorney Docket No.: F2099-7037WO Domain]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [TH Domain]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide located in the [Ast2 Domain]. In an embodiment, the 2’,5’-linked nucleotide is located at the nucleotide position 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of the TREM. In an embodiment, the 2’,5’-linked nucleotide is located at the nucleotide position 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or 75 of the TREM. In an embodiment, the TREM contains two or more 2’,5’-linked nucleotides. In an embodiment, the TREM contains two 2’,5’-linked nucleotides. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [ACH Domain]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [DH Domain]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [DH Domain] and the [TH Domain]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [TH Domain]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located in the [ASt Domain 1]. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located between nucleotide positions 30 and 40. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located between nucleotide positions 12 and 25. In an embodiment, the TREM contains two 2’,5’-linked nucleotides each independently located between nucleotide positions 30 and 40. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at nucleotide positions 31 and 32. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 31 and 34. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 31 and 35. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 31 and 37. In an embodiment, the TREM contains two 2’,5’-linked nucleotides located at the nucleotide positions 32 and 34. Attorney Docket No.: F2099-7037WO A TREM may comprise three 2’,5’-linked nucleotides. In an embodiment, the TREM comprises three 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].. In an embodiment, the TREM contains three 2’,5’-linked nucleotides each independently located between nucleotide positions 9 and 60. A TREM may comprise four 2’,5’-linked nucleotides. In an embodiment, the TREM comprises four 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. A TREM may comprise five 2’,5’-linked nucleotides. In an embodiment, the TREM comprises five 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2].A TREM may comprise six 2’,5’-linked nucleotides. In an embodiment, the TREM comprises six 2’,5’-linked nucleotides each independently located in the [ASt Domain 1], the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In an embodiment, the TREM contains a 2’,5’-linked nucleotide at nucleotide position 49. A TREM may contain several non-naturally occurring modifications in addition to 2’,5’- linked nucleotides. A TREM may comprise a non-naturally occurring modification in each of the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM comprises four 2’,5’-linked nucleotides independently located in the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2]. In an embodiment, the TREM comprises four 2’,5’-linked nucleotides independently located in at nucleotide positions 43, 45, 54, and 65. In an embodiment, the TREM comprises five 2’,5’-linked nucleotides, in addition to 24 2’-O-methyl modifications. In an embodiment, the TREM comprises five 2’,5’-linked nucleotides, in addition to 142’-fluoro modifications. In an embodiment, the TREM comprises five locked nucleic acids, in addition to one 2’-O-methoxyethyl modification. In an embodiment, the TREM comprises five locked nucleic acids, in addition to nine phosphorothioate modifications. In some embodiments, the TREM comprises five locked nucleic acids independently located in the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2], 242’-O-methyl modifications independently located in the Attorney Docket No.: F2099-7037WO [ASt Domain 1], [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2], 142’-fluoro modifications independently located in the [DH Domain], the [ACH Domain], the [VL Domain], the [TH Domain], and the [ASt Domain2], one 2’-O- methoxyethyl modification located in the [ACH Domain], and nine phosphorothioate modifications independently located in the [ASt Domain1], [DH Domain], the [ACH Domain], the [TH Domain], and the [ASt Domain2]. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.176. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.193. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.202. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.204. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.207. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.216. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.217. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.218. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.219. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.220. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.222. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.224. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.226. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.227. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.228. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.229. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.232. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.233. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.238. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.239. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.240. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.241. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.242. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.243. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.244. In some Attorney Docket No.: F2099-7037WO embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.245. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.246. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.247. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.248. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.249. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.250. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.251. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.252. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.253. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.254. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.255. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.256. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.257. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.258. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.259. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.260. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.262. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.263. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.264. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.265. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.266. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.267. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.269. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.270. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.272. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.273. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.274. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.275. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.276. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.277. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.278. In some Attorney Docket No.: F2099-7037WO embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.279. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.281. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.282. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.283. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.284. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.285. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.286. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.287. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.288. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.289. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.290. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.291. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.292. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.293. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.294. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.295. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.296. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.297. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.298. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.299. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.301. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.302. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.304. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.305. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.307. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.309. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.310. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.311. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.312. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.313. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.315. In some Attorney Docket No.: F2099-7037WO embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.316. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.317. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.318. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.320. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.321. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.322. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.323. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.324. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.325. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.326. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.327. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.328. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.329. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.330. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.331. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.332. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.333. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.334. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.335. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.336. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.337. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.338. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.339. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.340. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.341. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.342. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.343. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.346. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.357. In some embodiments, the TREM containing a 2’,5’-linked nucleotide is TREM No.361. TREMS CONTAINING ADDITIONAL MODIFICATIONS Attorney Docket No.: F2099-7037WO Described herein are TREMs comprising a locked nucleic acid moiety or a 2’,5’-linked nucleotide, and at least one additional non-naturally occurring modifications. In one aspect, the TREM comprises a locked nucleic acid moiety and a 2’,5’-linked nucleotide. In an embodiment, the TREM comprises a locked nucleic acid moiety and a 2’,5’-linked nucleotide, as well as an additional non-naturally occurring modification. In some embodiments, the TREM comprises a single LNA moiety or 2’,5’-linked nucleotide. In some embodiments, the TREM comprises a plurality of LNA moieties and 2’,5’- linked nucleotides. In some embodiments, the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 LNA moieties or 2’,5’-linked nucleotides. In some embodiments, the TREM comprises at least one LNA moiety and at least one 2’,5’-linked nucleotide. In some embodiments, the TREM comprises one LNA moiety and one 2’,5’-linked nucleotide. In some embodiments, the TREM comprising an LNA moiety and a 2’,5’-linked nucleotide is a TREM having any one of the SEQ ID Nos: 1060, 1062, 1112, 1135, 1153-1158, 1161, 1163, 1166-1170, 1175, 1200, 1207, 1357, 1364, 1378-1393, and 1397-1398. In some embodiments, the TREM comprises two LNA moieties and at least one 2’,5’- linked nucleotide. In some embodiments, the TREM comprises two LNA moieties and one 2’,5’- linked nucleotide. In some embodiments, the TREM comprises two LNA moieties and two 2’,5’- linked nucleotides. In some embodiments, the TREM comprises two LNA moieties and three 2’,5’-linked nucleotides. In some embodiments, the TREM comprises two LNA moieties and four 2’,5’-linked nucleotides. In some embodiments, the TREM comprises three LNA moieties and at least one 2’,5’- linked nucleotide. In some embodiments, the TREM comprises three LNA moieties and one 2’,5’-linked nucleotide. In some embodiments, the TREM comprises three LNA moieties and two 2’,5’-linked nucleotides. In some embodiments, the TREM comprises three LNA moieties and three 2’,5’-linked nucleotides. In some embodiments, the TREM comprises four LNA moieties and at least one 2’,5’- linked nucleotide. In some embodiments, the TREM comprises four LNA moieties and one 2’,5’-linked nucleotide. In some embodiments, the TREM comprises four LNA moieties and two 2’,5’-linked nucleotides. In some embodiments, the TREM comprising both LNA moieties and 2’,5’-linked nucleotides also comprises at least one additional non-naturally occurring modification (e.g., 2’- Attorney Docket No.: F2099-7037WO ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’- deoxy, or phosphorothiorate modifications). In some embodiments, the TREM comprising an LNA moiety and a 2’,5’-linked nucleotide comprises at least 2, 3, 4, 5, 6, 7, 8.9, 10, or more additional non-naturally occurring modifications (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’- halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications). In some embodiments, the TREM comprises one LNA moiety, one 2’,5’-linked nucleotide, and at least one 5-methylcytosine. In some embodiments, the TREM comprises one LNA moiety, one 2’,5’-linked nucleotide, and one 5-methylcytosine. In some embodiments, the TREM comprises one LNA moiety, one 2’,5’-linked nucleotide, and at least one 2’-moe. In one aspect, the present disclosure is a method of treating a subject having a disease or disorder associated with a PTC comprising administering to the subject a TREM, TREM core fragment, or TREM fragment described herein (e.g., a TREM described herein), thereby treating the subject having the disease or disorder. In an embodiment, the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease. In an embodiment, the TREM comprising an LNA moiety or a 2’,5’-linked nucleotide comprises a sequence corresponding to the nucleotide sequence of SEQ ID NO.625. In an embodiment, the TREM comprising an LNA moiety or a 2’,5’-linked nucleotide comprises a sequence corresponding to the nucleotide sequence of SEQ ID NO.1416. TREM, TREM core fragment and TREM fragment fusions In an embodiment, a TREM, a TREM core fragment or a TREM fragment disclosed herein comprises an additional moiety, e.g., a fusion moiety. In an embodiment, the fusion moiety can be used for purification, to alter folding of the TREM, TREM core fragment or TREM fragment, or as a targeting moiety. In an embodiment, the fusion moiety can comprise a tag, a linker, can be cleavable or can include a binding site for an enzyme. In an embodiment, the fusion moiety can be disposed at the N terminal of the TREM or at the C terminal of the TREM, TREM core fragment or TREM fragment. In an embodiment, the fusion moiety can be encoded by the same or different nucleic acid molecule that encodes the TREM, TREM core fragment or TREM fragment. Attorney Docket No.: F2099-7037WO TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises a consensus sequence provided herein. In an embodiment, a TREM disclosed herein comprises a consensus sequence of Formula I ZZZ, wherein ZZZ indicates any of the twenty amino acids and Formula I corresponds to all species. In an embodiment, a TREM disclosed herein comprises a consensus sequence of Formula II ZZZ, wherein ZZZ indicates any of the twenty amino acids and Formula II corresponds to mammals. In an embodiment, a TREM disclosed herein comprises a consensus sequence of Formula III ZZZ, wherein ZZZ indicates any of the twenty amino acids and Formula III corresponds to humans. In an embodiment, ZZZ indicates any of the twenty amino acids: alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, methionine, leucine, lysine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine. In an embodiment, a TREM disclosed herein comprises a property selected from the following: a) under physiological conditions residue R0 forms a linker region, e.g., a Linker 1 region; b) under physiological conditions residues R1-R2-R3-R4 -R5-R6-R7 and residues R65-R66- R67-R68-R69-R70-R71 form a stem region, e.g., an AStD stem region; c) under physiological conditions residues R8-R9 forms a linker region, e.g., a Linker 2 region; d) under physiological conditions residues -R10-R11-R12-R13-R14 R15-R16-R17-R18-R19-R20- R21-R22-R23-R24-R25-R26-R27-R28 form a stem-loop region, e.g., a D arm Region; e) under physiological conditions residue -R29 forms a linker region, e.g., a Linker 3 Region; f) under physiological conditions residues -R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40- R41-R42-R43-R44-R45-R46 form a stem-loop region, e.g., an AC arm region; g) under physiological conditions residue -[R47]x comprises a variable region, e.g., as described herein; Attorney Docket No.: F2099-7037WO h) under physiological conditions residues -R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58- R59-R60-R61-R62-R63-R64 form a stem-loop region, e.g., a T arm Region; or i) under physiological conditions residue R72 forms a linker region, e.g., a Linker 4 region. Alanine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula IALA (SEQ ID NO: 562), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72, wherein R is a ribonucleotide residue and the consensus for Ala is: R^= absent; R^^, R^^=are independently A or absent; R^^= A, C, G or absent; R^, R^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently N or absent; R^^, R^^, R^^= are independently A, C, U or absent; R^, R^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, G or absent; R^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, G, U or absent; R^^, R^^= are independently A, U or absent; R^^, R^^=are independently C or absent; R^, R^= are independently C, G or absent; R^^, R^^, R^^, R^^, R^^= are independently C, G, U or absent; R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently C, U or absent; R^^, R^^, R^^= are independently G or absent; R^= G, U or absent; R^, R^^, R^^= are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1- 28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70-271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Attorney Docket No.: F2099-7037WO In an embodiment, a TREM disclosed herein comprises the sequence of Formula IIALA (SEQ ID NO: 563), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Ala is: R^, R18= are absent; R^^, R^^, R^^=are independently A or absent; R^^, R^^, R^^= are independently A, C, G or absent; R^^, R^^, R^^, R^^= are independently N or absent; R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, C, U or absent; R^, R^, R^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, G or absent; R^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, G, U or absent; R^, R^^, R^^= are independently A, U or absent; R^^, R^^, R^^, R^^= are independently C or absent; R^= C, G or absent; R^^, R^^, R^^= are independently C, G, U or absent; R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently C, U or absent; R^, R^^, R^^, R^^, R^^, R^^= are independently G or absent; R^, R^, R^^= are independently G, U or absent; R^^, R^^= are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, Attorney Docket No.: F2099-7037WO x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula IIIALA (SEQ ID NO: 564), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Ala is: R^, R18= are absent; R^^, R^^, R^^, R^^=are independently A or absent; R^^, R^^, R^^= are independently A, C, G or absent; R^^, R^^, R^^= are independently N or absent; R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, C, U or absent; R^, R^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, G or absent; R^, R^^, R^^, R^^, R^^, R^^, R^^= are independently A, G, U or absent; R^, R^^= are independently A, U or absent; R^^, R^^, R^^, R^^, R^^= are independently C or absent; R^^, R^^, R^^= are independently C, G, U or absent; R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^, R^^= are independently C, U or absent; R^, R^, R^, R^, R^^, R^^, R^^, R^^, R^^= are independently G or absent; R^^, R^^= are independently G, U or absent; R^, R^^, R^^, R^^= are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- Attorney Docket No.: F2099-7037WO 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Arginine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I ARG (SEQ ID NO: 565), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Arg is: R^^=A or absent; R^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^= C or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^=are independently G or absent; R^,R^^=are independently G,U or absent; R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; Attorney Docket No.: F2099-7037WO wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II ARG (SEQ ID NO: 566), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Arg is: R18= absent; R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^=are independently C or absent; R^^,R^^=are independently C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; Attorney Docket No.: F2099-7037WO R^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III ARG (SEQ ID NO: 567), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Arg is: R18=is absent; R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^,R^,R^^=are independently A,C,G or absent; R^,R^,R^^,R^^=are independently N or absent; R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; Attorney Docket No.: F2099-7037WO R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^,R^^=are independently C,G or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^,R^^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Asparagine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I ASN (SEQ ID NO: 568), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Asn is: R^,R^^=are absent; R^^= A or absent; R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^= are independently N or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^= C or absent; R^^= C,G or absent; R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II ASN (SEQ ID NO: 569), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Asn is: R0,R18=are absent Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^,R^^,R^^=are independently A,C,G or absent; R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^,R^^,R^^=are independently C or absent; R^^,R^^=are independently C,G or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III ASN (SEQ ID NO: 570), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Asn is: R0,R18=are absent R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^,R^^,R^^=are independently A,C,G or absent; R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^,R^^,R^^=are independently C or absent; R^^,R^^=are independently C,G or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Attorney Docket No.: F2099-7037WO Aspartate TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I ASP (SEQ ID NO: 571), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Asp is: R0=absent R^^,R^^=are independently A,C or absent; R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^=are independently A,G,U or absent; R^^= C or absent; R^^,R^^=are independently C,G or absent; R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^= G,U or absent; R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, Attorney Docket No.: F2099-7037WO x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II ASP (SEQ ID NO: 572), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Asp is: R0,R17,R18,R23=are independently absent; R^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^^,R^^=are independently A,C,G or absent; R^,R^,R^^=are independently N or absent; R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^=are independently A,U or absent; R^^,R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^=are independently C,G or absent; R^,R^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, Attorney Docket No.: F2099-7037WO x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III ASP (SEQ ID NO: 573), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Asp is: R0,R17,R18,R23=are absent R^,R^^,R^^,R^^,R^^=are independently A or absent; R^,R^^,R^^=are independently A,C or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^^= C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^^,R^^=are independently G,U or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- Attorney Docket No.: F2099-7037WO 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Cysteine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I CYS (SEQ ID NO: 574), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Cys is: R0 =absent R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^,R^^,R^^=are independently N or absent; R^^= A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^=are independently C or absent; R^= C, G or absent; R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^^,R^^=are independently U or absent; Attorney Docket No.: F2099-7037WO [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II CYS (SEQ ID NO: 575), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Cys is: R0,R18,R23=are absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^,R^^=are independently A,C,G or absent; R^^= A,C,G,U or absent; R^^,R^^=are independently A,C,U or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^,R^^=are independently C,G or absent; R^^= C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; Attorney Docket No.: F2099-7037WO R^,R^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; R^^= U, or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III CYS (SEQ ID NO: 576), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Cys is: R0,R18,R23=are absent R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^^= A,C,G or absent; R^^= N or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^^=are independently C,G or absent; Attorney Docket No.: F2099-7037WO R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Glutamine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I GLN (SEQ ID NO: 577), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Gln is: R^,R^^=are absent; R^^,R^^,R^^=are independently A or absent; R^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R ^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^,R^^=are independently A,G,U or absent; Attorney Docket No.: F2099-7037WO R^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II GLN (SEQ ID NO: 578), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Gln is: R0,R18,R23=are absent R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; Attorney Docket No.: F2099-7037WO R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III GLN (SEQ ID NO: 579), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Gln is: R0,R18,R23=are absent R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^=are independently A,C,U or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^,R^^,R^^=are independently C,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Glutamate TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I GLU (SEQ ID NO: 580), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Glu is: R^=absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^ ^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^= C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^= G or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II GLU (SEQ ID NO: 581), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Glu is: Attorney Docket No.: F2099-7037WO R0,R18,R23=are absent R^^,R^^=are independently A or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^= C or absent; R^^= C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III GLU (SEQ ID NO: 582), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Glu is: R0,R17,R18,R23=are absent R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^= A,C,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^=are independently A,G,U or absent; R^^,R^^,R^^,R^^=are independently A,U or absent; R^,R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C,G or absent; R^,R^^,R^^,R^^,R^^=are independently C,G,U o rabsent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^=are independently G or absent; R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), Attorney Docket No.: F2099-7037WO provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Glycine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I GLY (SEQ ID NO: 583), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Gly is: R^=absent; R^^= A or absent; R^,R^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R ^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^= A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^,R^^=are independently C or absent; R^^= C,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, Attorney Docket No.: F2099-7037WO x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II GLY (SEQ ID NO: 584), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Gly is: R0,R18,R23=are absent R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^,R^^=are independently A,C,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,G,U or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^=are independently C,G or absent; R^,R^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^=are independently G or absent; R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; Attorney Docket No.: F2099-7037WO wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III GLY (SEQ ID NO: 585), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Gly is: R0,R18,R23=are absent R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^,R^^,R^^=are independently A,C,G or absent; R^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^=are independently C,G or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; Attorney Docket No.: F2099-7037WO R^,R^^,R^^,R^^,R^^=are independently G or absent; R^^,R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Histidine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I HIS (SEQ ID NO: 586), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for His is: R^^=absent; R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^=are independently A,G,U or absent; Attorney Docket No.: F2099-7037WO R^^,R^^=are independently C or absent; R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^= G,U or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II HIS (SEQ ID NO: 587), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for His is: R0,R17,R18,R23=are absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^= A,C,U or absent; R^,R^^,R^^,R^^=are independently A,G or absent; R^^= A,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; Attorney Docket No.: F2099-7037WO R^^= C,G or absent; R^= C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III HIS (SEQ ID NO: 588), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for His is: R0,R17,R18,R23=are absent R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^= A,C or absent; R^,R^^,R^^,R^^=are independently A,G or absent; R^^= A,U or absent; Attorney Docket No.: F2099-7037WO R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Isoleucine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I ILE (SEQ ID NO: 589), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Ile is: R^^=absent; R^^,R^^,R^^,R^^=are independently A or absent; R^,R^^=are independently A,C,G or absent; R0,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R ^^,R^^=are independently N or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; Attorney Docket No.: F2099-7037WO R^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^= C or absent; R^,R^^,R^^=are independently C,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II ILE (SEQ ID NO: 590), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Ile is: R0,R18,R23=are absent R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^^,R^^,R^^=are independently N or absent; Attorney Docket No.: F2099-7037WO R^^= A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^^,R^^=are independently C,G or absent; R^= C,G,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III ILE (SEQ ID NO: 591), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Ile is: Attorney Docket No.: F2099-7037WO R0,R18,R23=are absent R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^= A,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^,R^^=are independently C,G or absent; R^^,R^^=are independently C,G,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Methionine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I MET (SEQ ID NO: 592), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Met is: R^,R^^=are absent; R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^ ^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^= C or absent; R^^,R^^=are independently C,G or absent; R^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^=are independently C,U or absent; R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Attorney Docket No.: F2099-7037WO In an embodiment, a TREM disclosed herein comprises the sequence of Formula II MET (SEQ ID NO: 593), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Met is: R0,R18,R22,R23=are absent R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^,R^^,R^^=are independently A,C or absent; R^,R^^,R^^=are independently A,C,G or absent; R^= N or absent; R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^=A,U or absent; R^,R^^,R^^=are independently C or absent; R^^,R^^,R^^,R^^=are independently C,G or absent; R^,R^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- Attorney Docket No.: F2099-7037WO 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III MET (SEQ ID NO: 594), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Met is: R0,R18,R22,R23=are absent R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^,R^^=are independently A,C or absent; R^,R^^=are independently A,C,G or absent; R^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^=are independently A,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^,R^^=are independently C,G or absent; R^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^=are independently U or absent; [R47] x = N or absent; Attorney Docket No.: F2099-7037WO wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Leucine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I LEU (SEQ ID NO: 595), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Leu is: R^=absent; R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^=are independently C,U or absent; R^^= G or absent; R^^= G,U or absent; Attorney Docket No.: F2099-7037WO R^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II LEU (SEQ ID NO: 596), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Leu is: R0 =absent R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^= C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III LEU (SEQ ID NO: 597), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Leu is: R0 =absent R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^,R^^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; Attorney Docket No.: F2099-7037WO R^^= A,U or absent; R^^= C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Lysine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I LYS (SEQ ID NO: 598), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Lys is: R0 =absent R^^= A or absent; R^^,R^^=are independently A,C or absent; R^^,R^^,R^^=are independently A,C,G or absent; Attorney Docket No.: F2099-7037WO R^,R^,R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^=are independently C,G or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II LYS (SEQ ID NO: 599), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Lys is: R0,R18,R23=are absent R^^= A or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^=are independently A,C or absent; R^,R^=are independently A,C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^,R^^=are independently C,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^=G,U or absent; R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III LYS (SEQ ID NO: 600), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 Attorney Docket No.: F2099-7037WO wherein R is a ribonucleotide residue and the consensus for Lys is: R0,R18,R23=absent R^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^,R^,R^^=are independently A,C,G or absent; R^^,R^^,R^^=are independently N or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^,R^^=are independently A,U or absent; R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^,R^^=are independently C,G or absent; R^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Attorney Docket No.: F2099-7037WO Phenylalanine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I PHE (SEQ ID NO: 601), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Phe is: R0,R23=are absent R^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^=are independently C or absent; R^,R^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^= G,U or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- Attorney Docket No.: F2099-7037WO 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II PHE (SEQ ID NO: 602), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Phe is: R0,R18,R23=absent R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^,R^^=are independently A,C,G or absent; R^^= A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^= A,G,U or absent; R^,R^,R^^=are independently A,U or absent; R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^,R^^=are independently C,G or absent; R^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; Attorney Docket No.: F2099-7037WO wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III PHE (SEQ ID NO: 603), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Phe is: R0,R18,R22,R23=absent R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^= A,C,G or absent; R^,R^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^=are independently A,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^^= C,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; Attorney Docket No.: F2099-7037WO wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Proline TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I PRO (SEQ ID NO: 604), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Pro is: R0 =absent R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently C or absent; R^,R^,R^^=are independently C,G or absent; R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^= are independently G,U or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II PRO (SEQ ID NO: 605), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Pro is: R0,R17,R18,R22,R23=absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C,G or absent; R^^=N or absent; R^^= A, C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^=are independently C,G or absent; R^^= C,G,U or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^^,R^^=are independently G,U or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III PRO (SEQ ID NO: 606), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Pro is: R0,R17,R18,R22,R23=absent R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C,U or absent; R^^,R^^,R^^=are independently A,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^=are independently C,G or absent; R^^= C,G,U or absent; Attorney Docket No.: F2099-7037WO R^^= C,U or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Serine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I SER (SEQ ID NO: 607), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Ser is: R^=absent; R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^,R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^= A,C,U or absent; R^^,R^^,R^^,R^^=are independently A,G or absent; Attorney Docket No.: F2099-7037WO R^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^= C or absent; R^^= C,G or absent; R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^^= G,U or absent; R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II SER (SEQ ID NO: 608), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Ser is: R0,R23=absent R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^=are independently C,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III SER (SEQ ID NO: 609), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 Attorney Docket No.: F2099-7037WO wherein R is a ribonucleotide residue and the consensus for Ser is: R0,R23=absent R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^=are independently C,G or absent; R^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Attorney Docket No.: F2099-7037WO Threonine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I THR (SEQ ID NO: 610), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Thr is: R0,R23=absent R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R ^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^=are independently C,G or absent; R^,R^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, Attorney Docket No.: F2099-7037WO x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II THR (SEQ ID NO: 611), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Thr is: R0,R18,R23=absent R^^,R^^,R^^=are independently A or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^= C or absent; R^^,R^^=are independently C,G or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- Attorney Docket No.: F2099-7037WO 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III THR (SEQ ID NO: 612), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Thr is: R0,R18,R23=absent R^^,R^^,R^^,R^^=are independently A or absent; R^^= A,C or absent; R^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^= C or absent; R^,R^^,R^^,R^^=are independently C,G or absent; R^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^^= G,U or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; Attorney Docket No.: F2099-7037WO wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Tryptophan TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I TRP (SEQ ID NO: 613), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Trp is: R^= absent; R^^,R^^,R^^,R^^=are independently A or absent; R^,R^,R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; Attorney Docket No.: F2099-7037WO [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II TRP (SEQ ID NO: 614), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Trp is: R0,R18,R22,R23=absent R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^,R^,R^^,R^^,R^^=are independently A,C or absent; R^,R^^=are independently A,C,G or absent; R^^= A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^= A,G,U or absent; R^^= A,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; Attorney Docket No.: F2099-7037WO R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III TRP (SEQ ID NO: 615), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Trp is: R0,R18,R22,R23=absent R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^,R^,R^^,R^^,R^^=are independently A,C or absent; R^,R^^=are independently A,C,G or absent; R^^= A,C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^^= A,G,U or absent; R^^= A,U or absent; Attorney Docket No.: F2099-7037WO R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,G or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G,U or absent; R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Tyrosine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I TYR (SEQ ID NO: 616), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Tyr is: R0 =absent R^^,R^^,R^^=are independently A or absent; R^^,R^^,R^^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^, R^^,R^^,R^^,R^^=are independently N or absent; Attorney Docket No.: F2099-7037WO R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^=are independently A,G,U or absent; R^^,R^^=are independently A,U or absent; R^^,R^^=are independently C or absent; R^^= C,G or absent; R^,R^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^=are independently G or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II TYR (SEQ ID NO: 617), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Tyr is: R0,R18,R23=absent R^,R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; Attorney Docket No.: F2099-7037WO R^^,R^^=are independently A,C or absent; R^^= A,C,G or absent; R^^= N or absent; R^^= A,C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^=are independently A,U or absent; R^^,R^^,R^^,R^^=are independently C or absent; R^^,R^^,R^^,R^^,R^^=are independently C,G or absent; R^,R^,R^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^,R^^,R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III TYR (SEQ ID NO: 618), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- Attorney Docket No.: F2099-7037WO R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Tyr is: R0,R18,R23=absent R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^=are independently A,C or absent; R^^= A,C,G or absent; R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^=are independently A,G,U or absent; R^^,R^^,R^^=are independently A,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C or absent; R^,R^^,R^^,R^^,R^^=are independently C,G or absent; R^,R^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently G or absent; R^,R^,R^^,R^^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Attorney Docket No.: F2099-7037WO Valine TREM Consensus sequence In an embodiment, a TREM disclosed herein comprises the sequence of Formula I VAL (SEQ ID NO: 619), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R 23 -R 24 -R 25 -R 26 -R 27 -R 28 -R 29 -R 30 -R 31 -R 32 -R 33 -R 34 -R 35 -R 36 -R 37 -R 38 -R 39 -R 40 -R 41 -R 42 - R 43 - R 44 -R 45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Val is: R^,R^^=absent; R^^,R^^,R^^=are independently A or absent; R^,R^^=are independently A,C,G or absent; R^,R^,R^,R^,R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^ ^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^=are independently A,C,U or absent; R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^,R^^,R^^=are independently A,G,U or absent; R^^= C or absent; R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^= G or absent; R^^= G ,U or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, Attorney Docket No.: F2099-7037WO x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula II VAL (SEQ ID NO: 620), R0- R1- R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Val is: R0,R18,R23=absent; R^^,R^^,R^^=are independently A or absent; R^^,R^^,R^^=are independently A,C,G or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently N or absent; R^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^= C or absent; R^,R^^=are independently C,G or absent; R^,R^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^= G or absent; R^,R^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, Attorney Docket No.: F2099-7037WO x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. In an embodiment, a TREM disclosed herein comprises the sequence of Formula III VAL (SEQ ID NO: 621), R0- R1-R2- R3-R4 -R5-R6-R7-R8-R9-R10-R11-R12-R13-R14-R15-R16-R17-R18-R19-R20-R21-R22- R23-R24-R25-R26-R27-R28-R29-R30-R31-R32-R33-R34-R35-R36-R37-R38-R39-R40-R41-R42- R43- R44-R45- R46- [R47]x-R48-R49-R50-R51-R52-R53-R54-R55-R56-R57-R58-R59-R60-R61-R62-R63-R64-R65-R66-R67- R68-R69-R70-R71-R72 wherein R is a ribonucleotide residue and the consensus for Val is: R0,R18,R23=absent R^^,R^^,R^^,R^^,R^^=are independently A or absent; R^^,R^^,R^^=are independently A,C,G or absent; R^^,R^^,R^^=are independently N or absent; R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,C,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently A,G,U or absent; R^^= A,U or absent; R^^= C or absent; R^,R^^=are independently C,G or absent; R^,R^,R^^,R^^,R^^,R^^=are independently C,G,U or absent; R^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^,R^^=are independently C,U or absent; R^^,R^^,R^^=are independently G or absent; R^,R^,R^^,R^^=are independently G,U or absent; R^,R^^,R^^,R^^,R^^=are independently U or absent; [R47] x = N or absent; Attorney Docket No.: F2099-7037WO wherein, e.g., x=1-271 (e.g., x=1-250, x=1-225, x=1-200, x=1-175, x=1-150, x=1-125, x=1-100, x=1-75, x=1-50, x=1-40, x=1-30, x=1-29, x=1-28, x=1-27, x=1-26, x=1-25, x=1-24, x=1-23, x=1-22, x=1-21, x=1-20, x=1-19, x=1-18, x=1-17, x=1-16, x=1-15, x=1-14, x=1-13, x=1-12, x=1-11, x=1-10, x=10-271, x=20-271, x=30-271, x=40-271, x=50-271, x=60-271, x=70- 271, x=80-271, x=100-271, x=125-271, x=150-271, x=175-271, x=200-271, x=225-271, x=1, x=2, x=3, x=4, x=5, x=6, x=7, x=8, x=9, x=10, x=11, x=12, x=13, x=14, x=15, x=16, x=17, x=18, x=19, x=20, x=21, x=22, x=23, x=24, x=25, x=26, x=27, x=28, x=29, x=30, x=40, x=50, x=60, x=70, x=80, x=90, x=100, x=110, x=125, x=150, x=175, x=200, x=225, x=250, or x=271), provided that the TREM has one or both of the following properties: no more than 15% of the residues are N; or no more than 20 residues are absent. Variable region consensus sequence In an embodiment, a TREM disclosed herein comprises a variable region at position R47. In an embodiment, the variable region is 1-271 ribonucleotides in length (e.g.1-250, 1-225, 1- 200, 1-175, 1-150, 1-125, 1-100, 1-75, 1-50, 1-40, 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 10-271, 20-271, 30- 271, 40-271, 50-271, 60-271, 70-271, 80-271, 100-271, 125-271, 150-271, 175-271, 200-271, 225-271, 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, 30, 40, 50, 60, 70, 80, 90, 100, 110, 125, 150, 175, 200, 225, 250, or 271 ribonucleotides). In an embodiment, the variable region comprises any one, all or a combination of Adenine, Cytosine, Guanine or Uracil. In an embodiment, the variable region comprises a ribonucleic acid (RNA) sequence encoded by a deoxyribonucleic acid (DNA) sequence disclosed in Table 4, e.g., any one of SEQ ID NOs: 452-561 disclosed in Table 4. Table 4: Exemplary variable region sequences. SEQ ID NO SEQUENCE
Figure imgf000207_0001
Attorney Docket No.: F2099-7037WO 461 CAGGCGGGTTCTGCCCGCGC 462 CATACCTGCAAGGGTATC
Figure imgf000208_0001
Attorney Docket No.: F2099-7037WO 504 TCTTAGCAATAAGGT 505 TCTTGTAGGAGTTC
Figure imgf000209_0001
Attorney Docket No.: F2099-7037WO 96 547 TGTTCGTAAGGACTT 97 548 TTCACAGAAATGTGTC
Figure imgf000210_0001
Corresponding Nucleotide Positions To determine if a selected nucleotide position in a candidate sequence corresponds to a selected position in a reference sequence (e.g., SEQ ID NO: 622, SEQ ID NO: 623, SEQ ID NO: 624), one or more of the following Evaluations is performed. Evaluation A: 1.The candidate sequence is aligned with each of the consensus sequences in Tables 9 and 10. The consensus sequence(s) having the most positions aligned (and which has at least 60% of the positions of the candidate sequence aligned) is selected. The alignment is performed as is follows. The candidate sequence and an isodecoder consensus sequence from Tables 10A-10B are aligned based on a global pairwise alignment calculated with the Needleman–Wunsch algorithm when run with match scores from Table 11, a mismatch penalty of -1, a gap opening penalty of -1, and a gap extension penalty of -0.5, and no penalty for end gaps. The alignment with the highest overall alignment score is then used to determine the percent similarity between the candidate and the consensus sequence by counting the number of matched positions in the alignment, dividing it by the larger of the number of non- N bases in the candidate sequence or the consensus sequence, and multiplying the result by 100. In cases where multiple alignments (of the candidate and a single consensus sequence) tie for the same score, the percent similarity is the largest percent similarity calculated from the tied alignments. This process is repeated for the candidate sequence with each of the remaining Attorney Docket No.: F2099-7037WO isodecoder consensus sequences in Tables 10A-10B, and the alignment resulting in the greatest percent similarity is selected. If this alignment has a percent similarity equal to or greater than 60%, it is considered a valid alignment and used to relate positions in the candidate sequence to those in the consensus sequence, otherwise the candidate sequence is considered to have not aligned to any of the isodecoder consensus sequences. If there is a tie at this point, all tied consensus sequences are taken forward to step 2 in the analysis. 2. Using the selected consensus sequence(s) from step 1, one determines the consensus sequence position number that aligns with the selected position (e.g., a modified position) in the candidate sequence. One then assigns the position number of the aligned position in the consensus sequence to the selected position in the candidate sequence, in other words, the selected position in the candidate sequence is numbered according to the numbering of the consensus sequence. If there were tied consensus sequences from step one, and they give different position numbers in this step 2, then all such position numbers are taken forward to step 5. 3. The reference sequence is aligned with the consensus sequence chosen in step 1. The alignment is performed as described in step 1. 4. From the alignment in step 3, one determines the consensus sequence position number that aligns with the selected position (e.g., a modified position) in the reference sequence. One then assigns the position number of the aligned position in the consensus sequence to the selected position in the reference sequence, in other words, the selected position in the reference sequence is numbered according to the numbering of the consensus sequence. If there is a tie at this point, all tied consensus sequences are taken forward to step 5 in the analysis. 5. If a value for a position number determined for the reference sequence in step 2 is the same as the value for the position number determined for the candidate sequence in step 4, the positions are defined as corresponding. Evaluation B: The reference sequence (e.g., a TREM sequence described herein) and the candidate sequence are aligned with one another. The alignment is performed as follows. The reference sequence and the candidate sequence are aligned based on a global pairwise alignment calculated with the Needleman–Wunsch algorithm when run with match scores from Table 11, a mismatch penalty of -1, a gap opening penalty of -1, and a gap extension Attorney Docket No.: F2099-7037WO penalty of -0.5, and no penalty for end gaps. The alignment with the highest overall alignment score is then used to determine the percent similarity between the candidate and reference sequence by counting the number of matched based in the alignment, dividing it by the larger of the number of non-N bases in the candidate or reference sequence, and multiplying the result by 100. In cases where multiple alignments tie for the same score, the percent similarity is the largest percent similarity calculated from the tied alignments. If this alignment has a percent similarity equal to or greater than 60%, it is considered a valid alignment and used to relate positions in the candidate sequence to those in the reference sequence, otherwise the candidate sequence is considered to have not aligned to the reference sequence. If the selected nucleotide position in the reference sequence (e.g., a modified position) is paired with a selected nucleotide position (e.g., a modified position) in the candidate sequence, the positions are defined as corresponding. Evaluation C: The candidate sequence is assigned a nucleotide position number according to the comprehensive tRNA numbering system (CtNS), also referred to as the tRNAviz method (e.g., as described in Lin et al., Nucleic Acids Research, 47:W1, pages W542-W547, 2 July 2019), which serves as a global numbering system for tRNA molecules. The alignment is performed as follows. 1. The candidate sequence is assigned a nucleotide position according to the tRNAviz method. For a novel sequence not present in the tRNAviz database, the numbering for the closest sequence in the database is obtained. For example, if a TREM differs at any given nucleotide position from a sequence in the database, the numbering for the tRNA having the wildtype sequence at said given nucleotide position is used. 2. The reference sequence is assigned a nucleotide position according to the method described in 1. 3. If a value for a position number determined for the reference sequence in step 1 is the same as the value for the position number determined for the candidate sequence in step 2, the positions are defined as corresponding. If the selected position in the reference sequence and the candidate sequence are found to be corresponding in at least one of Evaluations A, B, and C, the positions correspond. For example, if two positions are found to be corresponding under Evaluation A, but do not Attorney Docket No.: F2099-7037WO correspond under Evaluation B or Evaluation C, the positions are defined as corresponding. Similarly, if two positions are found to be corresponding under Evaluation B, but do not correspond under Evaluation A or Evaluation C, the positions are defined as corresponding. In addition, if two positions are found to be corresponding under Evaluation C, but do not correspond under Evaluation A or Evaluation B, the positions are defined as corresponding The numbering given above is used for ease of presentation and does not imply a required sequence. If more than one Evaluation is performed, they can be performed in any order. Table 10A. Consensus sequence computationally generated for each isodecoder by aligning members of the isodecoder family SEQ ID Amino NO. Acid Anticodon Consensus sequence C C C C T G C
Figure imgf000213_0001
Attorney Docket No.: F2099-7037WO TCCTCGTTAGTATAGTGGTGAGTATCCCCGC CTGTCACGCGGGAGACCGGGGTTCGATTCCC C A C A G C G C G C T C T C G G G G
Figure imgf000214_0001
Attorney Docket No.: F2099-7037WO GGTAGCGTGGCCGAGCGGTCTAAGGCGCTG GATTTAGGCTCCAGTCTCTTCGGNGGCGTGG C T T C C C G G G G C T
Figure imgf000215_0001
Attorney Docket No.: F2099-7037WO GACCTCGTGGCGCAACGGTAGCGCGTCTGA CTCCAGATCAGAAGGTTGCGTGTTCAAATCA C G
Figure imgf000216_0001
Table 10B. Consensus sequence computationally generated for each isodecoder by aligning members of the isodecoder family SEQ ID Amino C C T T A A C A
Figure imgf000216_0002
Attorney Docket No.: F2099-7037WO GGCTCTGTGGCGCAATGGATNAGCGCATTGG ACTTCTAATTCAAAGGTTGCGGGTTCGAGTC C A T T G G A
Figure imgf000217_0001
Attorney Docket No.: F2099-7037WO GTCAGGATGGCCGAGCGGTCTAAGGCGCTGC GTTCAGGTCGCAGTCTCCCCTGGAGGCGTGG A G A C A C A T G T G T A A
Figure imgf000218_0001
Attorney Docket No.: F2099-7037WO GGCNCTGTGGCTNAGTNGGNTAAAGCGCCGG TCTCGTAAACCNGGAGATCNTGGGTTCGAAT G A G C G C G T C
Figure imgf000219_0001
Table 11: Score values alignment Candidate Reference Match
Figure imgf000219_0002
Attorney Docket No.: F2099-7037WO Premature termination codons (PTC) and ORFs comprising PTCs Mutations underlie many diseases. For example, a point mutation in the open reading frame (ORF) of a gene which creates a premature stop codon (PTC) can result in altered expression and/or activity of a polypeptide encoded by the gene. Table 15 provides single mutations in codons encoding amino acids which can result in a stop codon. In an embodiment, a PTC disclosed herein comprises a mutation disclosed in Table 15. In an embodiment, the codon having the first sequence or the PTC comprises a mutation disclosed in Table 15. In an embodiment, the non-mutated, e.g., wildtype, codon sequence of the codon having the first sequence or the PTC is an original codon sequence provided in Table 15 and the amino acid corresponding to the non-mutated codon is an original AA provided in Table 15. In an embodiment, the TREM, TREM core fragment or TREM fragment recognizes a stop codon and mediates incorporation of the original AA provided in Table 15 at the position of the stop codon. In an embodiment, the TREM, TREM core fragment or TREM fragment recognizes a stop codon and mediates incorporation of an amino acid belonging to the same group as the original AA, e.g., as provided in Table 16. Other genetic abnormalities, such as insertions and/or deletions can also result in a PTC in an ORF. Table 15. Select amino acids and stop codons Original AA Original codon One mutation to stop d
Figure imgf000220_0001
Attorney Docket No.: F2099-7037WO ARG CGA UGA GLY GGA UGA
Figure imgf000221_0001
Group Amino acid Nonpolar, aliphatic R group leucine
Figure imgf000221_0002
Disclosed herein, inter alia, are endogenous ORFs comprising a codon having a first sequence, e.g., a mutation, e.g., a PTC. An ORF having a PTC, e.g., as described herein, can be present, or part of in any gene. As an example, the ORF can be present or be part of any gene in the human genome. In an embodiment, a PTC disclosed herein is present in a gene disclosed in any one of Tables 17, 18, or 20. Exemplary genes having ORFs comprising a PTC are provided in Table 3. Table 17: Exemplary genes with ORFs having a PTC A2ML1 ARFGEF1 CACNA1G CNOT1 DLG4
Figure imgf000221_0003
Attorney Docket No.: F2099-7037WO ACAD8 ASCC3 CARS2 COL9A1 DPH1 ACADL ASH1L CCDC140 COQ4 DPYD-AS1
Figure imgf000222_0001
Attorney Docket No.: F2099-7037WO FIBP GTPBP3 KCNMA1 MARS1 NCAPH2 FLAD1 HACE1 KCNQ5 MARS2 NCF2
Figure imgf000223_0001
Attorney Docket No.: F2099-7037WO P4HA1 PRDM12 RRM2B SLC18A3 SYN2 PAK6 PREPL RS1 SLC20A2 SYNJ1
Figure imgf000224_0001
Attorney Docket No.: F2099-7037WO TXNRD2 ZAP70 UBA2 ZBTB20
Figure imgf000225_0001
NAGLU, DMD, GAA, RP1, RP2, ABCA4, PCDH15, REP1, GLA, MUT, TP53, and ATM. In an embodiment, the PTC is present within the FVIII gene and comprises an R mutation, e.g., an R2228X mutation. In an embodiment, the PTC is present within the FIX gene and comprises an R mutation, e.g., an R29X mutation, an R116X mutation, an R248X mutation, an R252X mutation, an R333X mutation, and/or an R338X mutation. In an embodiment, the PTC is present within the CFTR gene and comprises an R mutation, e.g., an R553X mutation. In an embodiment, the PTC is present within the MeCP2 gene and comprises an R mutation, e.g., an R168X mutation. In an embodiment, the PTC is present within the NAGLU gene and comprises an R mutation, e.g., an R626X mutation. In an embodiment, t the PTC is present within the DMD gene and comprises an R mutation, e.g., an R3881X mutation. In an embodiment, the PTC is present within the GAA gene and comprises an R mutation, e.g., an R854X mutation. In an embodiment, the PTC is present within the RP1 gene and comprises an R mutation, e.g., an R667X mutation. In an embodiment, the PTC is present within the RP2 gene and comprises an R mutation, e.g., an R120X mutation. In an embodiment, the PTC is present within the ABCA4 gene and comprises an R mutation, e.g., an R2030X mutation. In an embodiment, the PTC is present within the PCD gene and comprises an R mutation, e.g., an R245X mutation. In an embodiment, the PTC is present within the REP1 gene and comprises an R mutation, e.g., an R270X mutation. In an embodiment, the PTC is a mutation in the GLA gene, e.g., an R220X mutation and/or an R227X mutation. In an embodiment, the PTC is present within the MUT gene and comprises an R mutation, e.g., an R228X mutation, an R403X mutation, an R467X mutation, and/or an R727X mutation. In an embodiment, the PTC is present within the TP53 gene and comprises an R mutation, e.g., an R578X mutation. In an embodiment the PTC is present within the ATM gene and comprises an R mutation, e.g., an R35X mutation. Attorney Docket No.: F2099-7037WO Diseases or disorders associated with a PTC A TREM composition disclosed herein can be used treat a disorder or disease associated with a PTC, e.g., as described herein. Exemplary diseases or disorders associated with a PTC are listed in Tables 18, 19, and 20. In an embodiment, the subject has a disease or disorder provided in any one of Tables 4- 6. In an embodiment, the cell is associated with, e.g., is obtained from a subject who has, a disorder or disease listed in any one of Tables 18-20. For example, the disorder or disease can be chosen from the left column of Table 18. As another example, the disorder or disease is chosen from the left column of Table 18 and, in embodiments the PTC is in a gene chosen from the right column of Table 18, e.g., any one of the genes provided in the right column of Table 18. In some embodiments, the PTC is in a gene corresponding to the disorder or disease provided in the left column of Table 18. As a further non-limiting example, the PTC can be at a position provided in Table 18. As another example, the disorder or symptom is chosen from a disorder or disease provided in Table 19. As yet another exmaple, the disorder or symptom is chosen from a disorder or disease provided in Table 20. In an embodiment, the disorder or symptom is chosen from a disorder or disease provided in Table 20 and, in embodiments, the PTC is in any gene provided in Table 20. In an embodiment, the disorder or symptom is chosen from a disorder or disease provided in Table 20 and the PTC is in a corresponding gene provided in Table 20, e.g., a gene corresponding to the disease or disorder. In an embodiment, the disorder or symptom is chosen from a disorder or disease provided in Table 20 and the PTC is not in a gene provided in Table 20. In an embodiment of any of the methods disclosed herein, the PTC is at any position within the ORF of the gene, e.g., upstream of the naturally occurring stop codon. Table 18: Exemplary diseases or disorders Disease/disorder or protein Exemplary Point Mutation
Figure imgf000226_0001
Attorney Docket No.: F2099-7037WO Lynch syndrome NM 000251.2(MSH2):c.212-1G>A Breast-ovarian cancer, familial 1 NM 007294.3(BRCA1):c.963G>A er)
Figure imgf000227_0001
Attorney Docket No.: F2099-7037WO Angelman syndrome (p.Trp768Ter) SMPD1 NM 000543.4(SMPD1):c.168G>A
Figure imgf000228_0001
Attorney Docket No.: F2099-7037WO T to C point mutations Wilson disease NM_000053.3(ATP7B):c.3443T>C (p.Ile l l hr)
Figure imgf000229_0001
Attorney Docket No.: F2099-7037WO Table 19: Additional exemplary disorders 5q-syndrome Holt-Oram syndrome Adams-Oliver syndrome-1 Hypoparathyroidism d)
Figure imgf000230_0001
Attorney Docket No.: F2099-7037WO Table 20: Exemplary genes with ORFs comprising a PTC and exemplary disorders Gene Disease/Disorder AAAS Glucocorticoid deficiency with achalasia is ;
Figure imgf000231_0001
Attorney Docket No.: F2099-7037WO ACY1 Neurological conditions associated with aminoacylase-1 deficiency Severe combined immunodeficiency disease; Severe combined immunodeficiency -
Figure imgf000232_0001
Attorney Docket No.: F2099-7037WO ALG3 Congenital disorder of glycosylation ALMS1 Alstrom syndrome a-
Figure imgf000233_0001
Attorney Docket No.: F2099-7037WO APOC2, APOC4- APOC2 Padova apolipoprotein C-II deficiency; Apolipoprotein C2 deficiency c c e ; y;
Figure imgf000234_0001
Attorney Docket No.: F2099-7037WO ATP2C1 Familial benign pemphigus ATP6V0A2 ALG9 congenital disorder of glycosylation; Cutis laxa with osteodystrophy y; e
Figure imgf000235_0001
Attorney Docket No.: F2099-7037WO Bloom syndrome; Hereditary breast and ovarian cancer syndrome; Hereditary BLM cancer-predisposing syndrome t d e ; ; t d 2; al
Figure imgf000236_0001
Attorney Docket No.: F2099-7037WO C12orf65 Combined oxidative phosphorylation deficiency-7; Spastic paraplegia Neurodegeneration with brain iron accumulation-4; Autosomal recessive spastic t
Figure imgf000237_0001
Attorney Docket No.: F2099-7037WO Noonan syndrome-like disorder with or without juvenile myelomonocytic CBL leukemia C hi i b h l hi Cl i h i i r ; e
Figure imgf000238_0001
Attorney Docket No.: F2099-7037WO Congenital heart defects, intellectual developmental disorder, and dysmorphic CDK13 facial features 4; d ic ; ;
Figure imgf000239_0001
Attorney Docket No.: F2099-7037WO Bronchiectasis with or without elevated sweat chloride-1; CFTR-related disorders; CFTR, Congenital bilateral aplasia of vas deferens from CFTR mutation; Cystic fibrosis; ; r; a;
Figure imgf000240_0001
Attorney Docket No.: F2099-7037WO X-linked recessive nephrolithiasis; X-linked recessive nephrolithiasis with renal CLCN5 failure c
Figure imgf000241_0001
Attorney Docket No.: F2099-7037WO COL6A1 Bethlem myopathy-1 COL6A2 Bethlem myopathy-1; Ullrich congenital muscular dystrophy-1 - e
Figure imgf000242_0001
Attorney Docket No.: F2099-7037WO Cerebroretinal microangiopathy with calcifications and cysts; Cerebroretinal CTC1 microangiopathy with calcifications and cysts-1; Dyskeratosis congenita - ;
Figure imgf000243_0001
Attorney Docket No.: F2099-7037WO CYP4F22 Autosomal recessive congenital ichthyosis-5 CZ1P-ASNS, l
Figure imgf000244_0001
Attorney Docket No.: F2099-7037WO Non-syndromic male infertility due to sperm motility disorder; Spermatogenic DNAH1 failure-18 -
Figure imgf000245_0001
Attorney Docket No.: F2099-7037WO Arrhythmogenic right ventricular cardiomyopathy type 8; Arrhythmogenic right ventricular dysplasia, cardiomyopathy; Cardiac arrest; Dilated cardiomyopathy ith ll hi k td d t th i C di l h t ; 1; e g
Figure imgf000246_0001
Attorney Docket No.: F2099-7037WO Cardiovascular phenotype; X-linked Emery-Dreifuss muscular dystrophy-1; EMD Neuromuscular disease A l i i f IC A l d i h l i l l e
Figure imgf000247_0001
Attorney Docket No.: F2099-7037WO EYA1 Branchiootic syndrome; Melnick-Fraser syndrome; Rare genetic deafness Autosomal dominant deafness-10; Dilated cardiomyopathy-1J; Rare genetic d
Figure imgf000248_0001
Attorney Docket No.: F2099-7037WO FGF16 Metacarpal 4-5 fusion FGF3 Deafness with labyrinthine aplasia microtia and microdontia (LAMM) - - ; s
Figure imgf000249_0001
Attorney Docket No.: F2099-7037WO FUCA1 Fucosidosis FYCO1 Cataract-18 e
Figure imgf000250_0001
Attorney Docket No.: F2099-7037WO Bilateral conductive hearing impairment; Bilateral sensorineural hearing impairment; GJB2-related disorders; Dominant digenic deafness-GJB2/GJB3; Di i d f GJB2/GJB6 H i i i t At l d i t h ig al s
Figure imgf000251_0001
Attorney Docket No.: F2099-7037WO GRHL2 Autosomal dominant deafness-28 GRHL3 Van der Woude syndrome-2 d e
Figure imgf000252_0001
Attorney Docket No.: F2099-7037WO HMCN1 Age-related macular degeneration-1 HMGB3 Syndromic microphthalmia-13 al ;
Figure imgf000253_0001
Attorney Docket No.: F2099-7037WO Short Rib Polydactyly Syndrome; Short-rib thoracic dysplasia-16 with or without IFT52 polydactyly
Figure imgf000254_0001
Attorney Docket No.: F2099-7037WO History of neurodevelopmental disorder; Autosomal dominant mental retardation- KAT6A 32 ly rt
Figure imgf000255_0001
Attorney Docket No.: F2099-7037WO KISS1R Hypogonadotropic hypogonadism-8 without anosmia KIZ Retinitis pigmentosa-69 e
Figure imgf000256_0001
Attorney Docket No.: F2099-7037WO LHFPL5 Rare genetic deafness LHX3 Non-acquired combined pituitary hormone deficiency with spine abnormalities s
Figure imgf000257_0001
Attorney Docket No.: F2099-7037WO LTBP3 Dental anomalies and short stature LTBP4 Cutis laxa with severe pulmonary, gastrointestinal, and urinary abnormalities n, 3; e; s
Figure imgf000258_0001
Attorney Docket No.: F2099-7037WO MEFV Familial Mediterranean fever Hereditary cancer-predisposing syndrome; Somatic lipoma; Multiple endocrine er
Figure imgf000259_0001
Attorney Docket No.: F2099-7037WO Carcinoma of colon; Colon cancer; Glioblastoma; Hereditary cancer-predisposing syndrome; Hereditary nonpolyposis colon cancer; Lynch syndrome; Lynch d I Mli t t f di l Mli t t f i id e- al y- y
Figure imgf000260_0001
Attorney Docket No.: F2099-7037WO MYL2, LOC114827850 Familial hypertrophic cardiomyopathy-10 y ; er nd ; ne
Figure imgf000261_0001
Attorney Docket No.: F2099-7037WO Hypertrophic cardiomyopathy; Long QT syndrome; Primary dilated cardiomyopathy; Primary familial hypertrophic cardiomyopathy; Sudden NEBL l i d d th g ty
Figure imgf000262_0001
Attorney Docket No.: F2099-7037WO Abnormality of color vision; Cone-rod dystrophy; Enhanced s-cone syndrome; Horizontal nystagmus; NR2E3-related disorders; Retinal dystrophy; Retinitis NR2E3 i t Rtiiti i t 37 Vi l i i t e e
Figure imgf000263_0001
Attorney Docket No.: F2099-7037WO Basal cell carcinoma; Breast cancer; Cancer of the pancreas; Familial cancer of breast; Complementation group N Fanconi anemia; Generalized h i tti H dit b t d i d H dit - s; ng y e A
Figure imgf000264_0001
Attorney Docket No.: F2099-7037WO Deafness/enamel hypoplasia/nail defects; Zellweger peroxisome biogenesis disorder 1A; Peroxisome biogenesis disorder 1B; Peroxisome biogenesis disorders; PEX1 GATAD1 Z ll d t is e
Figure imgf000265_0001
Attorney Docket No.: F2099-7037WO PKD1, LOC105371049 Adult type polycystic kidney disease A l i l i kid di Pl i kid d l i
Figure imgf000266_0001
Attorney Docket No.: F2099-7037WO Global developmental delay; Speech apraxia; White-sutton syndrome; Dysmorphy; POGZ Intellectual deficiency ’ ; g - g
Figure imgf000267_0001
Attorney Docket No.: F2099-7037WO PRDM16 Left ventricular noncompaction-8 PRDM5 Brittle cornea syndrome-2 ; or n
Figure imgf000268_0001
Attorney Docket No.: F2099-7037WO PUF60 Verheij syndrome Apnea; Generalized hypotonia; Autosomal dominant intellectual disability-31;
Figure imgf000269_0001
Attorney Docket No.: F2099-7037WO REEP6 Retinitis pigmentosa-77 RELT Amelogenesis imperfecta type IIIC 0
Figure imgf000270_0001
Attorney Docket No.: F2099-7037WO Congenital microcephaly; Microcephaly, short stature, and polymicrogyria with or RTTN without seizures A lid l k i F ilil l l di d ih i d lid r 2;
Figure imgf000271_0001
Attorney Docket No.: F2099-7037WO Carney-Stratakis syndrome; Gastrointestinal stromal tumor; Hereditary paraganglioma-pheochromocytoma syndromes; Hereditary cancer-predisposing SDHB d P li 4 Ph h t
Figure imgf000272_0001
Attorney Docket No.: F2099-7037WO Charcot-Marie-Tooth disease type 4C; Charcot-Marie-Tooth disease type 4; Inborn genetic diseases; Mild mononeuropathy of the median nerve; SH3TC2-related SH3TC2 di d h c
Figure imgf000273_0001
Attorney Docket No.: F2099-7037WO SLC9A3, SLC9A3-AS1 Congenital secretory sodium diarrhea-8
Figure imgf000274_0001
Attorney Docket No.: F2099-7037WO SPRTN Ruijs-Aalfs syndrome SPTA1 Elliptocytosis-2; Hereditary pyropoikilocytosis et - y;
Figure imgf000275_0001
Attorney Docket No.: F2099-7037WO Acute rhabdomyolysis; Cardiac arrhythmia; Episodic flaccid weakness; Intellectual functioning disability; Seizures; Recurrent metabolic crises with TANGO2 hbd l i di hth i d d ti
Figure imgf000276_0001
Attorney Docket No.: F2099-7037WO TFAP2B Patent ductus arteriosus-2 TFR2 Hemochromatosis type 3
Figure imgf000277_0001
Attorney Docket No.: F2099-7037WO Absent epiphyses; Chronic lung disease; Cleft palate; Clubfoot; Coat hanger sign of ribs; Common variable immune deficiency; Common variable i dfii 2 H i tb I lbli A dfii 2 It titil ; c ni al
Figure imgf000278_0001
Attorney Docket No.: F2099-7037WO Retinitis pigmentosa and erythrocytic microcytosis; Sideroblastic anemia with B- TRNT1 cell immunodeficiency, periodic fevers, and developmental delay s
Figure imgf000279_0001
Attorney Docket No.: F2099-7037WO Broad-based gait; Cardiomyopathy; Cardiovascular phenotype; Congenital muscular dystrophy; Decreased patellar reflex; Delayed gross motor development; Diltd di th 1G Diltd di th 1S Ditl l k s; d y-
Figure imgf000280_0001
Attorney Docket No.: F2099-7037WO Autosomal recessive deafness-18; Rare genetic deafness; Retinal dystrophy; USH1C Retinitis pigmentosa; Usher syndrome type 1C; Usher syndrome type 1 ar d - us
Figure imgf000281_0001
Attorney Docket No.: F2099-7037WO Autosomal dominant nonsyndromic deafness-6; Diabetes mellitus and insipidus with optic atrophy and deafness; WFS1-related spectrum disorders; Wolfram-like WFS1 d D ; s;
Figure imgf000282_0001
In an embodiment, the disease or disorder associated with a PTC is a lysosomal storage disease (e.g., Fabry disease, Gaucher disease, or Niemann-Pick disease). In some embodiments, the disease or disorder associated with a PTC is Fabry disease. In an embodiment, upon administration of a TREM (e.g., a TREM described herein) to a cell or subject, the level of a GLA protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, Attorney Docket No.: F2099-7037WO 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a GLA protein in a healthy, non-Fabry disease fibroblast). In some embodiments, the disease or disorder associated with a PTC is a blood clotting disorder, e.g., Hemophilia B. In an embodiment, upon administration of a TREM (e.g., a TREM described herein) to a cell or subject, the level of a Factor IX (FIX) protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a FIX protein in a healthy, non-disease cell). In some embodiments, the disease or disorder associated with a PTC is an autosomal recessive disorder, such as neuronal ceroid lipofuscinosis type 2 (CNL2). In an embodiment, upon administration of a TREM (e.g., a TREM described herein) to a cell or subject, the level of a tripeptidyl peptidase 1 (TPP1) protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a TPP1 protein in a healthy, non-disease cell). In some embodiments, the disease or disorder associated with a PTC is a disease or disorder associated with hearing loss, such as Usher syndrome (e.g., Usher syndrome type 1F). In an embodiment, upon administration of a TREM (e.g., a TREM described herein) to a cell or subject, the level of a protocadherin 15 precursor (PCDH15) protein in the cell or subject is modulated, e.g., increased, by about 0.1%, 0.5%, 1%, 2%, 3%, 4% 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more, e.g., compared with a reference value (e.g., level of a PCDH15 protein in a healthy, non-disease cell).
Attorney Docket No.: F2099-7037WO In an embodiment, the disease or disorder associated with a PTC is a proliferative disease, such as a benign neoplasm or a cancer. In an embodiment, the proliferative disease is associated with a benign neoplasm. For example, a benign neoplasm may include adenoma, fibroma, hemangioma, tuberous sclerosis, and lipoma. All types of benign neoplasms disclosed herein or known in the art are contemplated as being within the scope of the disclosure. In an embodiment, the proliferative disease is a cancer. As used herein, the term “cancer” refers to a malignant neoplasm (Stedman’s Medical Dictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia, 1990). All types of cancers disclosed herein or known in the art are contemplated as being within the scope of the disclosure. Exemplary cancers include, but are not limited to, acoustic neuroma; adenocarcinoma; adrenal gland cancer; anal cancer; angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma); appendix cancer; benign monoclonal gammopathy; biliary cancer (e.g., cholangiocarcinoma); bladder cancer; breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast); brain cancer (e.g., meningioma, glioblastomas, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma); bronchus cancer; carcinoid tumor; cervical cancer (e.g., cervical adenocarcinoma); choriocarcinoma; chordoma; craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma); connective tissue cancer; epithelial carcinoma; ependymoma; endotheliosarcoma (e.g., Kaposi’s sarcoma, multiple idiopathic hemorrhagic sarcoma); endometrial cancer (e.g., uterine cancer, uterine sarcoma); esophageal cancer (e.g., adenocarcinoma of the esophagus, Barrett’s adenocarcinoma); Ewing’s sarcoma; eye cancer (e.g., intraocular melanoma, retinoblastoma); familiar hypereosinophilia; gall bladder cancer; gastric cancer (e.g., stomach adenocarcinoma); gastrointestinal stromal tumor (GIST); germ cell cancer; head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma), throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer), e.g., adenoid cystic carcinoma (ACC)); hematopoietic cancers (e.g., leukemia such as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL)); lymphoma such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell Attorney Docket No.: F2099-7037WO lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma), follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas (e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., Waldenström’s macroglobulinemia), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, and anaplastic large cell lymphoma); a mixture of one or more leukemia/lymphoma as described above; and multiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease); hemangioblastoma; hypopharynx cancer; inflammatory myofibroblastic tumors; immunocytic amyloidosis; kidney cancer (e.g., nephroblastoma a.k.a. Wilms’ tumor, renal cell carcinoma); liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma); lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung); leiomyosarcoma (LMS); mastocytosis (e.g., systemic mastocytosis); muscle cancer; myelodysplastic syndrome (MDS); mesothelioma; myeloproliferative disorder (MPD) (e.g., polycythemia vera (PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)); neuroblastoma; neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis); neuroendocrine cancer (e.g., gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor); osteosarcoma (e.g., bone cancer); ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma); papillary adenocarcinoma; pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), Islet cell tumors); penile cancer (e.g., Paget’s disease of the penis and scrotum); pinealoma; primitive neuroectodermal tumor (PNT); plasma cell neoplasia; paraneoplastic syndromes; intraepithelial neoplasms; prostate cancer (e.g., prostate adenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland cancer; skin cancer (e.g., Attorney Docket No.: F2099-7037WO squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)); small bowel cancer (e.g., appendix cancer); soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous gland carcinoma; small intestine cancer; sweat gland carcinoma; synovioma; testicular cancer (e.g., seminoma, testicular embryonal carcinoma); thyroid cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer); urethral cancer; vaginal cancer; and vulvar cancer (e.g., Paget’s disease of the vulva). In some embodiments, the cancer is a solid tumor, such as a sarcoma or a carcinoma (e.g., lung cancer, brain cancer, breast cancer, bladder cancer, prostate cancer, colon cancer, rectal cancer). In another aspect, the present disclosure features methods of treating a disease or disorder in a cell or subject by administration of a TREM (e.g., a TREM described herein) to the cell or subject. Exemplary diseases or disorders include hemophilias, aminoacidopathies, metal storage disorders, peroxisome biogenesis disorder, progressive rare lung disease, diseases related to lipid metabolism, diseases related to galactose metabolism, systemic organic acidemias, urea cycle disorders, cholestastis disorders, bilirubin metabolism disorders, lysososomal storage disorders, glycogen storage diseases, and oxalate metabolism disorders. In an embodiment, the disease or disorder is a hemophilia, e.g., hemophilia A or hemophilia B. In an embodiment, the disease or disorder is an aminoacidopathy, e.g., tyrosinemia type 1, tyrosinemia type 2, tyrosinemia type 3, maple syrup urine disease, alkaptonuria, or phenylketonuria. In an embodiment, the disease or disorder is a systemic organic acidemia, e.g., methylmalonic acidemia (MMUT), methylmalonic acidemia (non-MMUT), propionic acidemia type A, propionic acidemia type B, or isovaleric acidemia. In an embodiment, the disease or disorder is a urea cycle disorder, e.g, argininosuccinate lyase deficiency, argininosuccinate lyase deficiency-D, citrullinemia type 1, citrullinemia type 2, carbamoyl phosphate synthetase-D, ornithine transcarbamylase, arginemia, or hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome. In an embodiment, the disease or disorder is lysosomal storage disorder, e.g., mucopolysaccharidosis 1, mucopolysaccharidosis 2, Fabry disease, lysosomal acid lipas deficiency, Pompe disease, Gaucher disease, Niemann Pick A, or Niemann Pick B. In an embodiment, the disease or disorder is a bilirubin metabolism disorder, e.g., Crigler-Najjar syndrome. In an embodiment, the disease or disorder is a cholestastis disorder, e.g., progressive familial intrahepatic cholestasis Attorney Docket No.: F2099-7037WO (PFIC) type 1, PFIC type 2, or PFIC type 3. In an embodiment, the disease or disorder is a disease related to lipid metabolism, e.g., sitosterolemia (ABCG5) or sitosterolemia (ABCG8). In an embodiment, the disease or disorder is a glycogen storage disease, e.g., glycogen storage disease 1a, glycogen storage disease 1b, or glycogen storage disease 3a. In an embodiment, the disease or disorder is a metal storage disorder, e.g., Wilson disease or hereditary hemochromatosis. In an embodiment, the disease or disorder is a progressive rare lung disease, e.g., alpha-1 antitrypsin deficiency. In an embodiment, the disease or disorder is a peroxisome biogenesis disorder, e.g., PBD RCDP1. In an embodiment, the disease or disorder is an oxalate metabolism disorder, e.g, primary hyperoxaluria type 1, primary hyperoxaluria type 2, or primary hyperoxaluria type 3. In an embodiment, the disease or disorder is a congenital disorder related to Notch signaling, e.g., Alagille syndrome. In an embodiment, the disease or disorder is an amyloidosis, e.g., familial amyloid polyneuropathy. In one aspect, the present disclosure features a method of treating a disease or disorder in a subject, the method comprising administering to the subject a TREM comprising the nucleotide sequence of any one of the TREMS listed in FIG.1. In an embodiment, the disease or disorder is selected from a hemophilia, aminoacidopathy, metal storage disorder, peroxisome biogenesis disorder, progressive rare lung disease, disease related to lipid metabolism, disease related to galactose metabolism, systemic organic acidemia, urea cycle disorder, cholestastis disorder, bilirubin metabolism disorder, lysososomal storage disorder, glycogen storage disease, and oxalate metabolism disorder. In an embodiment, the TREM comprises the sequence of any one of SEQ ID NO: 622, 623, 624, 4249, 4386, 4834, 5630, 6707, 6749, 6947, or 8051, or a fragment or variant thereof. Method of making TREMs, TREM core fragments, and TREM fragments In vitro methods for synthesizing oligonucleotides are known in the art and can be used to make a TREM, a TREM core fragment or a TREM fragment disclosed herein. For example, a TREM, TREM core fragment or TREM fragment can be synthesized using solid state synthesis or liquid phase synthesis. In an embodiment, a TREM, a TREM core fragment or a TREM fragment made according to an in vitro synthesis method disclosed herein has a different modification profile Attorney Docket No.: F2099-7037WO compared to a TREM expressed and isolated from a cell, or compared to a naturally occurring tRNA. An exemplary method for making a modified TREM is provided in Example 1. The method provided in Example 1 can also be used to make a synthetic TREM core fragment or synthetic TREM fragment. Additional synthetic methods are disclosed in Hartsel SA et al., (2005) Oligonucleotide Synthesis, 033–050, the entire contents of which are hereby incorporated by reference. TREM composition In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, comprises a pharmaceutically acceptable excipient. Exemplary excipients include those provided in the FDA Inactive Ingredient Database (https://www.accessdata.fda.gov/scripts/cder/iig/index. Cfm). In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100 or 150 grams of TREM, TREM core fragment or TREM fragment. In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50 or 100 milligrams of TREM, TREM core fragment or TREM fragment. In an embodiment, a TREM composition, e.g., a TREM pharmaceutical composition, is at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95 or 99% dry weight TREMs, TREM core fragments or TREM fragments. In an embodiment, a TREM composition comprises at least 1 x 106 TREM molecules, at least 1 x 107 TREM molecules, at least 1 x 108 TREM molecules or at least 1 x 109 TREM molecules. In an embodiment, a TREM composition comprises at least 1 x 106 TREM core fragment molecules, at least 1 x 107 TREM core fragment molecules, at least 1 x 108 TREM core fragment molecules or at least 1 x 109 TREM core fragment molecules. In an embodiment, a TREM composition comprises at least 1 x 106 TREM fragment molecules, at least 1 x 107 TREM fragment molecules, at least 1 x 108 TREM fragment molecules or at least 1 x 109 TREM fragment molecules. Attorney Docket No.: F2099-7037WO In an embodiment, a TREM composition produced by any of the methods of making disclosed herein can be charged with an amino acid using an in vitro charging reaction as known in the art. In an embodiment, a TREM composition comprise one or more species of TREMs, TREM core fragments, or TREM fragments. In an embodiment, a TREM composition comprises a single species of TREM, TREM core fragment, or TREM fragment. In an embodiment, a TREM composition comprises a first TREM, TREM core fragment, or TREM fragment species and a second TREM, TREM core fragment, or TREM fragment species. In an embodiment, the TREM composition comprises X TREM, TREM core fragment, or TREM fragment species, wherein X=2, 3, 4, 5, 6, 7, 8, 9, or 10. In an embodiment, the TREM, TREM core fragment, or TREM fragment has at least 70, 75, 80, 85, 90, or 95, or has 100%, identity with a sequence encoded by a nucleic acid in Table 1. In an embodiment, the TREM comprises a consensus sequence provided herein. A TREM composition can be formulated as a liquid composition, as a lyophilized composition or as a frozen composition. In some embodiments, a TREM composition can be formulated to be suitable for pharmaceutical use, e.g., a pharmaceutical TREM composition. In an embodiment, a pharmaceutical TREM composition is substantially free of materials and/or reagents used to separate and/or purify a TREM, TREM core fragment, or TREM fragment. In some embodiments, a TREM composition can be formulated with water for injection. In some embodiments, a TREM composition formulated with water for injection is suitable for pharmaceutical use, e.g., comprises a pharmaceutical TREM composition. TREM characterization A TREM, TREM core fragment, or TREM fragment, or a TREM composition, e.g., a pharmaceutical TREM composition, produced by any of the methods disclosed herein can be assessed for a characteristic associated with the TREM, TREM core fragment, or TREM fragment or the TREM composition, such as purity, sterility, concentration, structure, or functional activity of the TREM, TREM core fragment, or TREM fragment. Any of the above- mentioned characteristics can be evaluated by providing a value for the characteristic, e.g., by evaluating or testing the TREM, TREM core fragment, or TREM fragment, or the TREM Attorney Docket No.: F2099-7037WO composition, or an intermediate in the production of the TREM composition. The value can also be compared with a standard or a reference value. Responsive to the evaluation, the TREM composition can be classified, e.g., as ready for release, meets production standard for human trials, complies with ISO standards, complies with cGMP standards, or complies with other pharmaceutical standards. Responsive to the evaluation, the TREM composition can be subjected to further processing, e.g., it can be divided into aliquots, e.g., into single or multi-dosage amounts, disposed in a container, e.g., an end-use vial, packaged, shipped, or put into commerce. In embodiments, in response to the evaluation, one or more of the characteristics can be modulated, processed or re-processed to optimize the TREM composition. For example, the TREM composition can be modulated, processed or re-processed to (i) increase the purity of the TREM composition; (ii) decrease the amount of fragments in the composition; (iii) decrease the amount of endotoxins in the composition; (iv) increase the in vitro translation activity of the composition; (v) increase the TREM concentration of the composition; or (vi) inactivate or remove any viral contaminants present in the composition, e.g., by reducing the pH of the composition or by filtration. In an embodiment, the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has a purity of at least 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, i.e., by mass. In an embodiment, the TREM (e.g., TREM composition or an intermediate in the production of the TREM composition) has less than 0.1%, 0,5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25% TREM fragments relative to full length TREMs. In an embodiment, the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has low levels or absence of endotoxins, e.g., a negative result as measured by the Limulus amebocyte lysate (LAL) test. In an embodiment, the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has in-vitro translation activity, e.g., as measured by an assay described in Examples 6-7. In an embodiment, the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has a TREM Attorney Docket No.: F2099-7037WO concentration of at least 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10 ng/mL, 50 ng/mL, 0.1 ug/mL, 0.5 ug/mL,1 ug/mL, 2 ug/mL, 5 ug/mL, 10 ug/mL, 20 ug/mL, 30 ug/mL, 40 ug/mL, 50 ug/mL, 60 ug/mL, 70 ug/mL, 80 ug/mL, 100 ug/mL, 200 ug/mL, 300 ug/mL, 500 ug/mL, 1000 ug/mL, 5000 ug/mL, 10,000 ug/mL, or 100,000 ug/mL. In an embodiment, the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) is sterile, e.g., the composition or preparation supports the growth of fewer than 100 viable microorganisms as tested under aseptic conditions, the composition or preparation meets the standard of USP <71>, and/or the composition or preparation meets the standard of USP <85>. In an embodiment, the TREM, TREM core fragment, or TREM fragment (e.g., TREM composition or an intermediate in the production of the TREM composition) has an undetectable level of viral contaminants, e.g., no viral contaminants. In an embodiment, any viral contaminant, e.g., residual virus, present in the composition is inactivated or removed. In an embodiment, any viral contaminant, e.g., residual virus, is inactivated, e.g., by reducing the pH of the composition. In an embodiment, any viral contaminant, e.g., residual virus, is removed, e.g., by filtration or other methods known in the field. TREM administration Any TREM composition or pharmaceutical composition described herein can be administered to a cell, tissue or subject, e.g., by direct administration to a cell, tissue and/or an organ in vitro, ex-vivo or in vivo. In-vivo administration may be via, e.g., by local, systemic and/or parenteral routes, for example intravenous, subcutaneous, intraperitoneal, intrathecal, intramuscular, ocular, nasal, urogenital, intradermal, dermal, enteral, intravitreal, intracerebral, intrathecal, or epidural. Vectors and Carriers In some embodiments the TREM, TREM core fragment, or TREM fragment or TREM composition described herein, is delivered to cells, e.g. mammalian cells or human cells, using a vector. The vector may be, e.g., a plasmid or a virus. In some embodiments, delivery is in vivo, in vitro, ex vivo, or in situ. In some embodiments, the virus is an adeno associated virus (AAV), a lentivirus, or an adenovirus. In some embodiments, the system or components of the system are Attorney Docket No.: F2099-7037WO delivered to cells with a viral-like particle or a virosome. In some embodiments, the delivery uses more than one virus, viral-like particle or virosome. Carriers A TREM, a TREM composition or a pharmaceutical TREM composition described herein may comprise, may be formulated with, or may be delivered in, a carrier. Viral vectors The carrier may be a viral vector (e.g., a viral vector comprising a sequence encoding a TREM, a TREM core fragment or a TREM fragment). The viral vector may be administered to a cell or to a subject (e.g., a human subject or animal model) to deliver a TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition. A viral vector may be systemically or locally administered (e.g., injected). Viral genomes provide a rich source of vectors that can be used for the efficient delivery of exogenous genes into a mammalian cell. Viral genomes are known in the art as useful vectors for delivery because the polynucleotides contained within such genomes are typically incorporated into the nuclear genome of a mammalian cell by generalized or specialized transduction. These processes occur as part of the natural viral replication cycle, and do not require added proteins or reagents in order to induce gene integration. Examples of viral vectors include a retrovirus (e.g., Retroviridae family viral vector), adenovirus (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvovirus (e.g., adeno-associated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e.g., influenza virus), rhabdovirus (e.g., rabies and vesicular stomatitis virus), paramyxovirus (e.g., measles and Sendai), positive strand RNA viruses, such as picornavirus and alphavirus, and double stranded DNA viruses including adenovirus, herpesvirus (e.g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus, replication deficient herpes virus), and poxvirus (e.g., vaccinia, modified vaccinia Ankara (MVA), fowlpox and canarypox). Other viruses include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, human papilloma virus, human foamy virus, and hepatitis virus, for example. Examples of retroviruses include: avian leukosis-sarcoma, avian C-type viruses, mammalian C- type, B-type viruses, D-type viruses, oncoretroviruses, HTLV-BLV group, lentivirus, alpharetrovirus, gammaretrovirus, spumavirus (Coffin, J. M., Retroviridae: The viruses and their replication, Virology (Third Edition) Lippincott-Raven, Philadelphia, 1996). Other examples include murine leukemia viruses, murine sarcoma viruses, mouse mammary tumor virus, bovine Attorney Docket No.: F2099-7037WO leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, Gibbon ape leukemia virus, Mason Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus and lentiviruses. Other examples of vectors are described, for example, in US Patent No.5,801,030, the teachings of which are incorporated herein by reference. In some embodiments the system or components of the system are delivered to cells with a viral-like particle or a virosome. Cell and vesicle-based carriers A TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition described herein can be administered to a cell in a vesicle or other membrane-based carrier. In embodiments, a TREM, a TREM core fragment or a TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein is administered in or via a cell, vesicle or other membrane-based carrier. In one embodiment, the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition can be formulated in liposomes or other similar vesicles. Liposomes are spherical vesicle structures composed of a uni- or multilamellar lipid bilayer surrounding internal aqueous compartments and a relatively impermeable outer lipophilic phospholipid bilayer. Liposomes may be anionic, neutral or cationic. Liposomes are biocompatible, nontoxic, can deliver both hydrophilic and lipophilic drug molecules, protect their cargo from degradation by plasma enzymes, and transport their load across biological membranes and the blood brain barrier (BBB) (see, e.g., Spuch and Navarro, Journal of Drug Delivery, vol.2011, Article ID 469679, 12 pages, 2011. doi:10.1155/2011/469679 for review). Vesicles can be made from several different types of lipids; however, phospholipids are most commonly used to generate liposomes as drug carriers. Methods for preparation of multilamellar vesicle lipids are known in the art (see for example U.S. Pat. No.6,693,086, the teachings of which relating to multilamellar vesicle lipid preparation are incorporated herein by reference). Although vesicle formation can be spontaneous when a lipid film is mixed with an aqueous solution, it can also be expedited by applying force in the form of shaking by using a homogenizer, sonicator, or an extrusion apparatus (see, e.g., Spuch and Navarro, Journal of Drug Delivery, vol.2011, Article ID 469679, 12 pages, 2011. doi:10.1155/2011/469679 for review). Extruded lipids can be prepared by extruding through filters of decreasing size, as described in Attorney Docket No.: F2099-7037WO Templeton et al., Nature Biotech, 15:647-652, 1997, the teachings of which relating to extruded lipid preparation are incorporated herein by reference. Lipid Nanoformulations/Lipid-based carriers Lipid nanoparticles are another example of a carrier that provides a biocompatible and biodegradable delivery system for the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition described herein. Nanostructured lipid carriers (NLCs) are modified solid lipid nanoparticles (SLNs) that retain the characteristics of the SLN, improve drug stability and loading capacity, and prevent drug leakage. Polymer nanoparticles (PNPs) are an important component of drug delivery. These nanoparticles can effectively direct drug delivery to specific targets and improve drug stability and controlled drug release. Lipid–polymer nanoparticles (PLNs), a new type of carrier that combines liposomes and polymers, may also be employed. These nanoparticles possess the complementary advantages of PNPs and liposomes. A PLN is composed of a core–shell structure; the polymer core provides a stable structure, and the phospholipid shell offers good biocompatibility. As such, the two components increase the drug encapsulation efficiency rate, facilitate surface modification, and prevent leakage of water-soluble drugs. For a review, see, e.g., Li et al.2017, Nanomaterials 7, 122; doi:10.3390/nano7060122. Exemplary lipid nanoparticles are disclosed in International Application PCT/US2014/053907, the entire contents of which are hereby incorporated by reference. For example, an LNP described in paragraphs [403-406] or [410-413] of PCT/US2014/053907 can be used as a carrier for the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition described herein. Additional exemplary lipid nanoparticles are disclosed in U.S. Patent 10,562,849 the entire contents of which are hereby incorporated by reference. For example, an LNP of formula (I) as described in columns 1-3 of U.S. Patent 10,562,849 can be used as a carrier for the TREM, TREM core fragment, TREM fragment, or TREM composition or pharmaceutical TREM composition described herein. Lipids that can be used in nanoparticle formations (e.g., lipid nanoparticles) include, for example those described in Table 4 of WO2019217941, which is incorporated by reference, e.g., a lipid-containing nanoparticle can comprise one or more of the lipids in Table 4 of Attorney Docket No.: F2099-7037WO WO2019217941. Lipid nanoparticles can include additional elements, such as polymers, such as the polymers described in Table 5 of WO2019217941, incorporated by reference. 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-methoxypoly ethylene glycol 2000)- 1 ,2-distearoyl-sn-glycero-3-phosphoethanolamine sodium salt, and those described in Table 2 of WO2019051289 (incorporated by reference), and combinations of the foregoing. In some embodiments, sterols that can be incorporated into lipid nanoparticles include one or more of cholesterol or cholesterol derivatives, such as those in W02009/127060 or US2010/0130588, which are incorporated by reference. Additional exemplary sterols include phytosterols, including those described in Eygeris et al (2020), incorporated herein by reference. Cationic Lipids (Positively Charged) and Ionizable Lipids 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. 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), 3β-[N-(N',N'- dimethylaminoethyl)carbamoyl) cholesterol (DC-chol), 1,2-dioleoyloxy-3-[trimethylammonio]- propane (DOTAP), 1,2-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)-1-(cis,cis-9,12- Attorney Docket No.: F2099-7037WO octadecadienoxy)propane (CLinDMA), 2-[5′-(cholest-5-en-3-beta-oxy)-3′-oxapentoxy)-3- dimethyl-1-(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, which is herein incorporated by reference in its entirety. In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises more than one cationic lipid. 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. 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, which is incorporated by reference. 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, , , Attorney Docket No.: F2099-7037WO ,
Figure imgf000297_0001
entirety). 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), which is incorporated herein by reference in its entirety. 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), (which is incorporated by reference herein in its entirety). 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; I, IA, 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; Attorney Docket No.: F2099-7037WO 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 WO2015/074085 (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; I-5 or I-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-O13 or 503- O13 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, all of which are incorporated herein by reference in their entirety. In some embodiments, the lipid-based carrier (or lipid nanoformulation) further includes biodegradable ionizable lipids, for instance, (9Z,l2Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,l2-dienoate, also called 3- ((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,l2Z)-octadeca-9,l2-dienoate). See, e.g., lipids of WO 2019/067992, WO 2017/173054, WO 2015/095340, and WO 2014/136086, which are incorporated herein by reference in their entirety. In some embodiments, the lipid particle comprises an ionizable lipid, a non-cationic lipid, a conjugated lipid that inhibits aggregation of particles, and a sterol. The amounts of these components can be varied independently and to achieve desired properties. For example, in some embodiments, the lipid nanoparticle comprises an ionizable lipid is in an amount from about 20 mol % to about 90 mol % of the total lipids (in other embodiments it may be 20-70% (mol), 30-60% (mol) or 40-50% (mol); about 50 mol % to about 90 mol % of the total lipid Attorney Docket No.: F2099-7037WO present in the lipid nanoparticle), a non-cationic lipid in an amount from about 5 mol % to about 30 mol % of the total lipids, a conjugated lipid in an amount from about 0.5 mol % to about 20 mol % of the total lipids, and a sterol in an amount from about 20 mol % to about 50 mol % of the total lipids. The ratio of total lipid to nucleic acid can be varied as desired. For example, the total lipid to nucleic acid (mass or weight) ratio can be from about 10: 1 to about 30: 1. In some embodiments, the lipid to nucleic acid 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 lipids and nucleic acid can be adjusted to provide a desired N/P ratio, for example, N/P ratio of 3, 4, 5, 6, 7, 8, 9, 10 or higher. Generally, the lipid nanoparticle formulation’s overall lipid content can range from about 5 mg/ml to about 30 mg/mL. Some non-limiting example of lipid compounds that may be used (e.g., in combination with other lipid components) to form lipid nanoparticles for the delivery of compositions described herein, e.g., nucleic acid (e.g., RNA) described herein includes, (i)
Figure imgf000299_0001
a TREM composition described herein to the liver and/or hepatocyte cells.
Figure imgf000299_0002
a TREM composition described herein to the liver and/or hepatocyte cells.
Attorney Docket No.: F2099-7037WO In some embodiments an LNP comprising Formula (iii) is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells. to deliver a TREM
Figure imgf000300_0001
composition described herein to the liver and/or hepatocyte cells.
Figure imgf000300_0002
is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells.
Attorney Docket No.: F2099-7037WO a TREM
Figure imgf000301_0001
Figure imgf000301_0002
In some embodiments an LNP comprising Formula (ix) is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells. alkylene, X3 is C(=O) or a 1
Figure imgf000301_0003
direct bond, R is H or Me, R3 is Ci-3 alkyl, R2 is Ci-3 alkyl, or R2 taken together with the nitrogen atom to which it is attached and 1-3 carbon atoms of X2 form a 4-, 5-, or 6-membered ring, or X1 is NR1, R1 and R2 taken together with the nitrogen atoms to which they are attached form a 5- or 6-membered ring, or R2 taken together with R3 and the nitrogen atom to which they are attached form a 5-, 6-, or 7-membered ring, Y1 is C2-12 alkylene, Y2 is selected from
Figure imgf000301_0004
(in either orientation), Attorney Docket No.: F2099-7037WO n is 0 to 3, R4 is Ci-15 alkyl, Z1 is Ci-6 alkylene or a direct bond, , (in either orientation) or absent, provided that if Z1 is a direct bond, Z2 is alkyl or C6-10
Figure imgf000302_0001
alkoxy, R6 is C5-9 alkyl or C6-10 alkoxy, W is methylene or a direct bond, and R7 is H or Me, or a salt thereof, provided that if R3 and R2 are C2 alkyls, X1 is O, X2 is linear C3 alkylene, X3 is C(=0), Y1 is linear Ce alkylene, (Y2)n-R4 is , R4 is linear C5 alkyl, Z1 is C2 alkylene, Z2 is absent, W is methylene, 6
Figure imgf000302_0002
and R are not Cx alkoxy. In some embodiments an LNP comprising Formula (xii) is used to deliver a TREM composition described herein to the liver and/or hepatocyte cells. TREM
Figure imgf000302_0003
composition described herein to the liver and/or hepatocyte cells.
Figure imgf000302_0004
In some embodiments an LNP comprises a compound of Formula (xiii) and a compound of Formula (xiv). Attorney Docket No.: F2099-7037WO a TREM
Figure imgf000303_0001
an LNP comprising a formulation of Formula (xvi) is used to
Figure imgf000303_0002
deliver a TREM composition described herein to the lung endothelial cells. (xviii) (a)
Figure imgf000303_0003
Attorney Docket No.: F2099-7037WO to form lipid nanoparticles for the delivery
Figure imgf000304_0001
of e.g., a herein is made by one of the following reactions: (a)
Figure imgf000304_0002
provided in an LNP that comprises an ionizable lipid. In some embodiments, the ionizable lipid is heptadecan-9-yl 8-((2-hydroxyethyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate (SM-102); e.g., as described in Example 1 of US9,867,888 (incorporated by reference herein in its entirety). In some embodiments, the ionizable lipid is 9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,12-dienoate (LP01), e.g., as synthesized in Example 13 of WO2015/095340 (incorporated by reference herein in its entirety). In some embodiments, the ionizable lipid is Di((Z)-non-2-en-1-yl) 9-((4-dimethylamino)- butanoyl)oxy)heptadecanedioate (L319), e.g. as synthesized in Example 7, 8, or 9 of US2012/0027803 (incorporated by reference herein in its entirety). In some embodiments, the ionizable lipid is 1,1’-((2-(4-(2-((2-(Bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl) amino)ethyl)piperazin-1-yl)ethyl)azanediyl)bis(dodecan-2-ol) (C12-200), e.g., as synthesized in Examples 14 and 16 of WO2010/053572 (incorporated by reference herein in its entirety). In some embodiments, the ionizable lipid is Imidazole cholesterol ester (ICE) lipid (3S, 10R, 13R, 17R)-10, 13-dimethyl-17- ((R)-6-methylheptan-2-yl)-2, 3, 4, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-lH- cyclopenta[a]phenanthren-3-yl 3-(1H-imidazol-4-yl)propanoate, e.g., Structure (I) from WO2020/106946 (incorporated by reference herein in its entirety). Attorney Docket No.: F2099-7037WO In some embodiments, an ionizable lipid may be a cationic lipid, an ionizable cationic lipid, 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. Exemplary cationic lipids include one or more amine group(s) which bear the positive charge. In some embodiments, the lipid particle comprises a cationic lipid in formulation with one or more of neutral lipids, ionizable amine-containing lipids, biodegradable alkyne lipids, steroids, phospholipids including polyunsaturated lipids, structural lipids (e.g., sterols), PEG, cholesterol and polymer conjugated lipids. In some embodiments, the cationic lipid may be an ionizable cationic lipid. An exemplary cationic lipid as disclosed herein may have an effective pKa over 6.0. In embodiments, a lipid nanoparticle may comprise a second cationic lipid having a different effective pKa (e.g., greater than the first effective pKa), than the first cationic lipid. A lipid nanoparticle may comprise between 40 and 60 mol percent of a cationic lipid, a neutral lipid, a steroid, a polymer conjugated lipid, and a therapeutic agent, e.g., a TREM described herein, encapsulated within or associated with the lipid nanoparticle. In some embodiments, the TREM is co-formulated with the cationic lipid. The TREM may be adsorbed to the surface of an LNP, e.g., an LNP comprising a cationic lipid. In some embodiments, the TREM may be encapsulated in an LNP, e.g., an LNP comprising a cationic lipid. In some embodiments, the lipid nanoparticle may comprise a targeting moiety, e.g., coated with a targeting agent. In embodiments, the LNP formulation is biodegradable. In some embodiments, a lipid nanoparticle comprising one or more lipid described herein, e.g., Formula (i), (ii), (ii), (vii) and/or (ix) encapsulates at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 92%, at least 95%, at least 97%, at least 98% or 100% of a TREM. Exemplary ionizable lipids that can be used in lipid nanoparticle formulations include, without limitation, those listed in Table 1 of WO2019051289, incorporated herein by reference. Additional exemplary lipids include, without limitation, one or more of the following formulae: X of US2016/0311759; I of US20150376115 or in US2016/0376224; I, II or III of US20160151284; I, IA, II, or IIA of US20170210967; I-c of US20150140070; A of US2013/0178541; I of US2013/0303587 or US2013/0123338; I of US2015/0141678; II, III, IV, or V of US2015/0239926; I of US2017/0119904; I or II of WO2017/117528; A of Attorney Docket No.: F2099-7037WO US2012/0149894; A of US2015/0057373; A of WO2013/116126; A of US2013/0090372; A of US2013/0274523; A of US2013/0274504; A of US2013/0053572; A of W02013/016058; A of W02012/162210; I of US2008/042973; I, II, III, or IV of US2012/01287670; I or II of US2014/0200257; I, II, or III of US2015/0203446; I or III of US2015/0005363; I, IA, IB, IC, ID, II, IIA, IIB, IIC, IID, or III-XXIV of US2014/0308304; of US2013/0338210; I, II, III, or IV of W02009/132131; A of US2012/01011478; I or XXXV of US2012/0027796; XIV or XVII of US2012/0058144; of US2013/0323269; I of US2011/0117125; I, II, or III of US2011/0256175; I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII of US2012/0202871; I, II, III, IV, V, VI, VII, VIII, X, XII, XIII, XIV, XV, or XVI of US2011/0076335; I or II of US2006/008378; I of US2013/0123338; I or X-A-Y-Z of US2015/0064242; XVI, XVII, or XVIII of US2013/0022649; I, II, or III of US2013/0116307; I, II, or III of US2013/0116307; I or II of US2010/0062967; I-X of US2013/0189351; I of US2014/0039032; V of US2018/0028664; I of US2016/0317458; I of US2013/0195920; 5, 6, or 10 of US10,221,127; III-3 of WO2018/081480; I-5 or I-8 of WO2020/081938; 18 or 25 of US9,867,888; A of US2019/0136231; II of WO2020/219876; 1 of US2012/0027803; OF-02 of US2019/0240349; 23 of US10,086,013; cKK-E12/A6 of Miao et al (2020); C12-200 of WO2010/053572; 7C1 of Dahlman et al (2017); 304-O13 or 503-O13 of Whitehead et al; TS-P4C2 of US9,708,628; I of WO2020/106946; I of WO2020/106946. In some embodiments, the ionizable lipid is MC3 (6Z,9Z,28Z,3 lZ)-heptatriaconta- 6,9,28,3 l-tetraen-l9-yl-4-(dimethylamino) butanoate (DLin-MC3-DMA or MC3), e.g., as described in Example 9 of WO2019051289A9 (incorporated by reference herein in its entirety). In some embodiments, the ionizable lipid is the lipid ATX-002, e.g., as described in Example 10 of WO2019051289A9 (incorporated by reference herein in its entirety). In some embodiments, the ionizable lipid is (l3Z,l6Z)-A,A-dimethyl-3- nonyldocosa-l3, l6-dien-l-amine (Compound 32), e.g., as described in Example 11 of WO2019051289A9 (incorporated by reference herein in its entirety). In some embodiments, the ionizable lipid is Compound 6 or Compound 22, e.g., as described in Example 12 of WO2019051289A9 (incorporated by reference herein in its entirety). Non-Cationic Lipids (e.g., Phospholipids) 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 Attorney Docket No.: F2099-7037WO replacement. In some embodiments, the non-cationic lipid is a negatively charged (anionic) lipid. 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-l-trans PE, 1-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), dielaidoyl- phosphatidylethanolamine (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, which is incorporated herein by reference. Such lipids include, in some embodiments, plant lipids found to improve liver transfection with mRNA (e.g., DGTS). In some embodiments, the lipid-based carrier (or lipid nanoformulation) may comprise a combination of distearoylphosphatidylcholine/cholesterol, Attorney Docket No.: F2099-7037WO dipalmitoylphosphatidylcholine/cholesterol, dimyrystoylphosphatidylcholine/cholesterol, 1,2- Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/cholesterol, or egg sphingomyelin/cholesterol. 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 non- cationic lipids are described in WO 2017/099823 or US 2018/0028664, which are incorporated herein by reference in their entirety. 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, which is incorporated herein by reference in its entirety. 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. 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). In some embodiments, the lipid-based carrier (or lipid nanoformulation) does not include any phospholipids. 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). 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 Attorney Docket No.: F2099-7037WO (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), monomethyl-phosphatidylethanolamine (such as 16-O-monomethyl PE), dimethyl- phosphatidylethanolamine (such as 16-O-dimethyl PE), l8-l-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), dielaidoyl- phosphatidylethanolamine (DEPE), lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidicacid,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, incorporated herein by reference. Such lipids include, in some embodiments, plant lipids found to improve liver transfection with mRNA (e.g., DGTS). Other examples of non-cationic lipids suitable for use in the lipid nanoparticles include, without limitation, nonphosphorous lipids such as, e.g., stearylamine, dodeeylamine, hexadecylamine, acetyl palmitate, glycerol ricinoleate, hexadecyl stereate, 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 non-cationic lipids are described in WO2017/099823 or US patent publication US2018/0028664, the contents of which is incorporated herein by reference in their entirety. In some embodiments, the non-cationic lipid is oleic acid or a compound of Formula I, II, or IV of US2018/0028664, incorporated herein by reference in its entirety. The non-cationic lipid can comprise, for example, 0-30% (mol) of the total lipid present in the lipid nanoparticle. In some embodiments, the non-cationic lipid content is 5-20% (mol) or 10-15% (mol) of the total lipid present in the lipid nanoparticle. In embodiments, the molar ratio of ionizable lipid to the 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). Attorney Docket No.: F2099-7037WO In some embodiments, the lipid nanoparticles do not comprise any phospholipids. Structural Lipids 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. 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. 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). 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, which are incorporated herein by reference in their entirety. Additional exemplary sterols include phytosterols, including those described in Eygeris et al. (2020), Nano Lett.2020;20(6):4543- 4549, incorporated herein by reference. 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)-buty1 ether. Exemplary cholesterol derivatives are described in WO 2009/127060 and US 2010/0130588, each of which is incorporated herein by reference in its entirety. 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. Attorney Docket No.: F2099-7037WO In some aspects, the lipid nanoparticle can further comprise a component, such as a sterol, to provide membrane integrity. One exemplary sterol that can be used in the lipid nanoparticle is cholesterol and derivatives thereof. Non-limiting examples of cholesterol derivatives include polar analogues such as 5a-choiestanol, 53-coprostanol, choiesteryl-(2,- hydroxy)-ethyl ether, choiesteryl-(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., choiesteryl-(4 ‘-hydroxy)-buty1 ether. Exemplary cholesterol derivatives are described in PCT publication W02009/127060 and US patent publication US2010/0130588, each of which is incorporated herein by reference in its entirety. In some embodiments, the component providing membrane integrity, such as a sterol, can comprise 0-50% (mol) (e.g., 0-10%, 10-20%, 20-30%, 30-40%, or 40-50%) of the total lipid present in the lipid nanoparticle. In some embodiments, such a component is 20-50% (mol) 30- 40% (mol) of the total lipid content of the lipid nanoparticle. Polymers and Polyethylene Glycol (PEG) - Lipids 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. 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, 1,2-Diacyl-sn-Glycero-3- Phosphoethanolamine-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); 1,2-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); 1,2-Diacyl-sn-Glycero-3- Phosphoethanolamine-N- [Methoxy(Polyethylene glycol)-5000] (mPEG 5000 PE); N-Acyl- Sphingosine-1- [Succinyl(Methoxy Polyethylene Glycol) 750] (mPEG 750 Ceramide); N-Acyl- Sphingosine-1- [Succinyl(Methoxy Polyethylene Glycol) 2000] (mPEG 2000 Ceramide); and N- Acyl- Sphingosine-1-[Succinyl(Methoxy Polyethylene Glycol) 5000] (mPEG 5000 Ceramide). In some Attorney Docket No.: F2099-7037WO embodiments, the PEG lipid is a polyethyleneglycol-diacylglycerol (i.e., polyethyleneglycol diacylglycerol (PEG-DAG), PEG-cholesterol, or PEG-DMB) conjugate. 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, which is incorporated herein by reference in its entirety). 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). 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 (C18). 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 Table 2 of WO 2019/051289 (which is herein incorporated by reference in its entirety), and combinations of the foregoing. 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, all of which are incorporated herein by reference in their entirety. 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, both of which are incorporated herein by reference in their entirety. 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 Attorney Docket No.: F2099-7037WO includes one of the following:
Figure imgf000313_0001
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. 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, which is incorporated herein by reference in its entirety. 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. 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). 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), Attorney Docket No.: F2099-7037WO which are incorporated herein by reference in their entirety. In some embodiments, the lipid nanoparticle can comprise a polyethylene glycol (PEG) or a conjugated lipid molecule. Generally, these are used to inhibit aggregation of lipid nanoparticles and/or provide steric stabilization. Exemplary conjugated lipids include, but are not limited to, PEG-lipid conjugates, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (such as ATTA-lipid conjugates), cationic-polymer lipid (CPL) conjugates, and mixtures thereof. In some embodiments, the conjugated lipid molecule is a PEG-lipid conjugate, for example, a (methoxy polyethylene glycol)-conjugated lipid. Exemplary PEG-lipid conjugates include, but are not limited to, 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)-l,2- distearoyl-sn-glycero-3-phosphoethanolamine sodium salt, or a mixture thereof. Additional exemplary PEG-lipid conjugates are described, for example, in US5,885,6l3, US6,287,59l, US2003/0077829, US2003/0077829, US2005/0175682, US2008/0020058, US2011/0117125, US2010/0130588, US2016/0376224, US2017/0119904, and US/099823, the contents of all of which are incorporated herein by reference in their entirety. In some embodiments, a PEG-lipid is a compound of Formula III, III-a-I, III-a-2, III-b-1, III-b-2, or V of US2018/0028664, the content of which is incorporated herein by reference in its entirety. In some embodiments, a PEG-lipid is of Formula II of US20150376115 or US2016/0376224, the content of both of which is incorporated herein by reference in its entirety. In some embodiments, the PEG-DAA conjugate can be, for example, PEG-dilauryloxypropyl, PEG- dimyristyloxypropyl, PEG- dipalmityloxypropyl, or PEG-distearyloxypropyl. The PEG-lipid can be one or more of PEG- DMG, PEG-dilaurylglycerol, PEG-dipalmitoylglycerol, PEG- disterylglycerol, PEG- dilaurylglycamide, PEG-dimyristylglycamide, PEG- dipalmitoylglycamide, PEG- disterylglycamide, PEG-cholesterol (l-[8’-(Cholest-5-en-3[beta]- oxy)carboxamido-3’,6’- dioxaoctanyl] carbamoyl-[omega]-methyl-poly(ethylene glycol), PEG- DMB (3,4- Ditetradecoxylbenzyl- [omega]-methyl-poly(ethylene glycol) ether), and 1,2- dimyristoyl-sn- glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]. In some embodiments, Attorney Docket No.: F2099-7037WO the PEG-lipid comprises PEG-DMG, 1,2- dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000]. In some embodiments, the PEG-lipid comprises a structure selected from:
Figure imgf000315_0001
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. Exemplary conjugated lipids, i.e., PEG-lipids, (POZ)-lipid conjugates, ATTA-lipid conjugates and cationic polymer-lipids are described in the PCT and LIS patent applications listed in Table 2 of WO2019051289A9, the contents of all of which are incorporated herein by reference in their entirety. In some embodiments, the PEG or the conjugated lipid can comprise 0-20% (mol) of the total lipid present in the lipid nanoparticle. In some embodiments, PEG or the conjugated lipid content is 0.5- 10% or 2-5% (mol) of the total lipid present in the lipid nanoparticle. Molar ratios of the ionizable lipid, non-cationic-lipid, sterol, and PEG/conjugated lipid can be varied as needed. For example, the lipid particle can comprise 30-70% ionizable lipid by mole or by total weight of the composition, 0-60% cholesterol by mole or by total weight of the composition, 0- 30% non-cationic-lipid by mole or by total weight of the composition and 1-10% conjugated lipid by mole or by total weight of the composition. Preferably, the composition comprises 30- 40% ionizable lipid by mole or by total weight of the composition, 40-50% cholesterol by mole Attorney Docket No.: F2099-7037WO or by total weight of the composition, and 10- 20% non-cationic-lipid by mole or by total weight of the composition. In some other embodiments, the composition is 50-75% ionizable lipid by mole or by total weight of the composition, 20-40% cholesterol by mole or by total weight of the composition, and 5 to 10% non-cationic-lipid, by mole or by total weight of the composition and 1-10% conjugated lipid by mole or by total weight of the composition. The composition may contain 60-70% ionizable lipid by mole or by total weight of the composition, 25-35% cholesterol by mole or by total weight of the composition, and 5-10% non-cationic-lipid by mole or by total weight of the composition. The composition may also contain up to 90% ionizable lipid by mole or by total weight of the composition and 2 to 15% non-cationic lipid by mole or by total weight of the composition. The formulation may also be a lipid nanoparticle formulation, for example comprising 8-30% ionizable lipid by mole or by total weight of the composition, 5- 30% non- cationic lipid by mole or by total weight of the composition, and 0-20% cholesterol by mole or by total weight of the composition; 4-25% ionizable lipid by mole or by total weight of the composition, 4-25% non-cationic lipid by mole or by total weight of the composition, 2 to 25% cholesterol by mole or by total weight of the composition, 10 to 35% conjugate lipid by mole or by total weight of the composition, and 5% cholesterol by mole or by total weight of the composition; or 2-30% ionizable lipid by mole or by total weight of the composition, 2-30% non-cationic lipid by mole or by total weight of the composition, 1 to 15% cholesterol by mole or by total weight of the composition, 2 to 35% conjugate lipid by mole or by total weight of the composition, and 1-20% cholesterol by mole or by total weight of the composition; or even up to 90% ionizable lipid by mole or by total weight of the composition and 2-10% non-cationic lipids by mole or by total weight of the composition, or even 100% cationic lipid by mole or by total weight of the composition. In some embodiments, the lipid particle formulation comprises ionizable lipid, phospholipid, cholesterol and a PEG-ylated lipid in a molar ratio of 50: 10:38.5: 1.5. In some other embodiments, the lipid particle formulation comprises ionizable lipid, cholesterol and a PEG-ylated lipid in a molar ratio of 60:38.5: 1.5. In some embodiments, the lipid particle comprises ionizable lipid, non-cationic lipid (e.g. phospholipid), a sterol (e.g., cholesterol) and a PEG-ylated lipid, where the molar ratio of lipids ranges from 20 to 70 mole percent for the ionizable lipid, with a target of 40-60, the mole percent of non-cationic lipid ranges from 0 to 30, with a target of 0 to 15, the mole percent of sterol Attorney Docket No.: F2099-7037WO ranges from 20 to 70, with a target of 30 to 50, and the mole percent of PEG-ylated lipid ranges from 1 to 6, with a target of 2 to 5. In some embodiments, the lipid particle comprises ionizable lipid / non-cationic- lipid / sterol / conjugated lipid at a molar ratio of 50: 10:38.5: 1.5. In an aspect, the disclosure provides a lipid nanoparticle formulation comprising phospholipids, lecithin, phosphatidylcholine and phosphatidylethanolamine. In some embodiments, one or more additional compounds can also be included. Those compounds can be administered separately, or the additional compounds can be included in the lipid nanoparticles of the invention. In other words, the lipid nanoparticles can contain other compounds in addition to the nucleic acid or at least a second nucleic acid, different than the first. Without limitations, other additional compounds can be selected from the group consisting of small or large organic or inorganic molecules, monosaccharides, disaccharides, trisaccharides, oligosaccharides, polysaccharides, peptides, proteins, peptide analogs and derivatives thereof, peptidomimetics, nucleic acids, nucleic acid analogs and derivatives, an extract made from biological materials, or any combinations thereof. In some embodiments, LNPs are directed to specific tissues by the addition of targeting domains. For example, biological ligands may be displayed on the surface of LNPs to enhance interaction with cells displaying cognate receptors, thus driving association with and cargo delivery to tissues wherein cells express the receptor. In some embodiments, the biological ligand may be a ligand that drives delivery to the liver, e.g., LNPs that display GalNAc result in delivery of nucleic acid cargo to hepatocytes that display asialoglycoprotein receptor (ASGPR). The work of Akinc et al. Mol Ther 18(7):1357-1364 (2010) teaches the conjugation of a trivalent GalNAc ligand to a PEG-lipid (GalNAc-PEG-DSG) to yield LNPs dependent on ASGPR for observable LNP cargo effect (see, e.g., FIG.6 of Akinc et al.2010, supra). Other ligand- displaying LNP formulations, e.g., incorporating folate, transferrin, or antibodies, are discussed in WO2017223135, which is incorporated herein by reference in its entirety, in addition to the references used therein, namely Kolhatkar et al., Curr Drug Discov Technol.20118:197-206; Musacchio and Torchilin, Front Biosci.201116:1388-1412; Yu et al., Mol Membr Biol.2010 27:286-298; Patil et al., Crit Rev Ther Drug Carrier Syst.200825:1-61 ; Benoit et al., Biomacromolecules.201112:2708-2714; Zhao et al., Expert Opin Drug Deliv.20085:309-319; Akinc et al., Mol Ther.201018:1357-1364; Srinivasan et al., Methods Mol Biol.2012820:105- Attorney Docket No.: F2099-7037WO 116; Ben-Arie et al., Methods Mol Biol.2012757:497-507; Peer 2010 J Control Release.20:63- 68; Peer et al., Proc Natl Acad Sci U S A.2007104:4095-4100; Kim et al., Methods Mol Biol. 2011721:339-353; Subramanya et al., Mol Ther.201018:2028-2037; Song et al., Nat Biotechnol.200523:709-717; Peer et al., Science.2008319:627-630; and Peer and Lieberman, Gene Ther.201118:1127-1133. In some embodiments, LNPs are selected for tissue-specific activity by the addition of a Selective ORgan Targeting (SORT) molecule to a formulation comprising traditional components, such as ionizable cationic lipids, amphipathic phospholipids, cholesterol and poly(ethylene glycol) (PEG) lipids. The teachings of Cheng et al. Nat Nanotechnol 15(4):313- 320 (2020) demonstrate that the addition of a supplemental “SORT” component precisely alters the in vivo RNA delivery profile and mediates tissue-specific (e.g., lungs, liver, spleen) gene delivery and editing as a function of the percentage and biophysical property of the SORT molecule. In some embodiments, the LNPs comprise biodegradable, ionizable lipids. In some embodiments, the LNPs comprise (9Z,l2Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,l2-dienoate, also called 3- ((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,l2Z)-octadeca-9,l2-dienoate) or another ionizable lipid. See, e.g, lipids of WO2019/067992, WO/2017/173054, WO2015/095340, and WO2014/136086, as well as references provided therein. In some embodiments, the term cationic and ionizable in the context of LNP lipids is interchangeable, e.g., wherein ionizable lipids are cationic depending on the pH. In some embodiments, the average LNP diameter of the LNP formulation may be between 10s of nm and 100s of nm, e.g., measured by dynamic light scattering (DLS). In some embodiments, the average LNP diameter of the LNP formulation may be from about 40 nm to about 150 nm, such as about 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm. In some embodiments, the average LNP diameter of the LNP formulation may be from about 50 nm to about 100 nm, from about 50 nm to about 90 nm, from about 50 nm to about 80 nm, from about 50 nm to about 70 nm, from about 50 nm to about 60 nm, from about 60 nm to about 100 nm, from about 60 nm to about 90 nm, from about 60 nm to about 80 nm, from about 60 nm to about 70 nm, from about 70 nm to about 100 nm, from about Attorney Docket No.: F2099-7037WO 70 nm to about 90 nm, from about 70 nm to about 80 nm, from about 80 nm to about 100 nm, from about 80 nm to about 90 nm, or from about 90 nm to about 100 nm. In some embodiments, the average LNP diameter of the LNP formulation may be from about 70 nm to about 100 nm. In a particular embodiment, the average LNP diameter of the LNP formulation may be about 80 nm. In some embodiments, the average LNP diameter of the LNP formulation may be about 100 nm. In some embodiments, the average LNP diameter of the LNP formulation ranges from about l mm to about 500 mm, from about 5 mm to about 200 mm, from about 10 mm to about 100 mm, from about 20 mm to about 80 mm, from about 25 mm to about 60 mm, from about 30 mm to about 55 mm, from about 35 mm to about 50 mm, or from about 38 mm to about 42 mm. A LNP may, in some instances, be relatively homogenous. A polydispersity index may be used to indicate the homogeneity of a LNP, e.g., the particle size distribution of the lipid nanoparticles. A small (e.g., less than 0.3) polydispersity index generally indicates a narrow particle size distribution. A LNP may have a polydispersity index from about 0 to about 0.25, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, or 0.25. In some embodiments, the polydispersity index of a LNP may be from about 0.10 to about 0.20. The zeta potential of a LNP may be used to indicate the electrokinetic potential of the composition. In some embodiments, the zeta potential may describe the surface charge of an LNP. Lipid nanoparticles with relatively low charges, positive or negative, are generally desirable, as more highly charged species may interact undesirably with cells, tissues, and other elements in the body. In some embodiments, the zeta potential of a LNP may be from about -10 mV to about +20 mV, from about -10 mV to about +15 mV, from about -10 mV to about +10 mV, from about -10 mV to about +5 mV, from about -10 mV to about 0 mV, from about -10 mV to about -5 mV, from about -5 mV to about +20 mV, from about -5 mV to about +15 mV, from about -5 mV to about +10 mV, from about -5 mV to about +5 mV, from about -5 mV to about 0 mV, from about 0 mV to about +20 mV, from about 0 mV to about +15 mV, from about 0 mV to about +10 mV, from about 0 mV to about +5 mV, from about +5 mV to about +20 mV, from about +5 mV to about +15 mV, or from about +5 mV to about +10 mV. The efficiency of encapsulation of a TREM describes the amount of TREM that is encapsulated or otherwise associated with a LNP after preparation, relative to the initial amount provided. The encapsulation efficiency is desirably high (e.g., close to 100%). The encapsulation Attorney Docket No.: F2099-7037WO efficiency may be measured, for example, by comparing the amount of TREM in a solution containing the lipid nanoparticle before and after breaking up the lipid nanoparticle with one or more organic solvents or detergents. An anion exchange resin may be used to measure the amount of free protein or nucleic acid (e.g., RNA) in a solution. Fluorescence may be used to measure the amount of free TREM in a solution. For the lipid nanoparticles described herein, the encapsulation efficiency of a TREM 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 80%. In some embodiments, the encapsulation efficiency may be at least 90%. In some embodiments, the encapsulation efficiency may be at least 95%. A LNP may optionally comprise one or more coatings. In some embodiments, a LNP may be formulated in a capsule, film, or table having a coating. A capsule, film, or tablet including a composition described herein may have any useful size, tensile strength, hardness or density. Additional exemplary lipids, formulations, methods, and characterization of LNPs are taught by WO2020061457, which is incorporated herein by reference in its entirety. In some embodiments, in vitro or ex vivo cell lipofections are performed using Lipofectamine MessengerMax (Thermo Fisher) or TransIT-mRNA Transfection Reagent (Mirus Bio). In certain embodiments, LNPs are formulated using the GenVoy_ILM ionizable lipid mix (Precision NanoSystems). In certain embodiments, LNPs are formulated using 2,2‐dilinoleyl‐4‐ dimethylaminoethyl‐[1,3]‐dioxolane (DLin‐KC2‐DMA) or dilinoleylmethyl‐4‐ dimethylaminobutyrate (DLin-MC3-DMA or MC3), the formulation and in vivo use of which are taught in Jayaraman et al. Angew Chem Int Ed Engl 51(34):8529-8533 (2012), incorporated herein by reference in its entirety. LNP formulations optimized for the delivery of CRISPR-Cas systems, e.g., Cas9-gRNA RNP, gRNA, Cas9 mRNA, are described in WO2019067992 and WO2019067910, both incorporated by reference. Additional specific LNP formulations useful for delivery of nucleic acids are described in US8158601 and US8168775, both incorporated by reference, which include formulations used in patisiran, sold under the name ONPATTRO. Attorney Docket No.: F2099-7037WO Exosomes can also be used as drug delivery vehicles for the TREM, TREM core fragment, TREM fragment, or TREM compositions or pharmaceutical TREM composition described herein. For a review, see Ha et al. July 2016. Acta Pharmaceutica Sinica B. Volume 6, Issue 4, Pages 287-296; https://doi.org/10.1016/j.apsb.2016.02.001. Ex vivo differentiated red blood cells can also be used as a carrier for a TREM, TREM core fragment, TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein. See, e.g., WO2015073587; WO2017123646; WO2017123644; WO2018102740; wO2016183482; WO2015153102; WO2018151829; WO2018009838; Shi et al.2014. Proc Natl Acad Sci USA.111(28): 10131–10136; US Patent 9,644,180; Huang et al. 2017. Nature Communications 8: 423; Shi et al.2014. Proc Natl Acad Sci USA.111(28): 10131–10136. Fusosome compositions, e.g., as described in WO2018208728, can also be used as carriers to deliver the TREM, TREM core fragment, TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein. Virosomes and virus-like particles (VLPs) can also be used as carriers to deliver a TREM, TREM core fragment, TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein to targeted cells. Plant nanovesicles, e.g., as described in WO2011097480A1, WO2013070324A1, or WO2017004526A1 can also be used as carriers to deliver the TREM, TREM core fragment, TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein. Percentages of Lipid Nanoformulation Components In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises one of more of the compounds described herein, 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 a TREM (e.g., a TREM described herein). 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 Attorney Docket No.: F2099-7037WO 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%. 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. In some embodiments, the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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 payload may be at least 70%. 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. In some embodiments, the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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 payload may be at least 70%. 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 Attorney Docket No.: F2099-7037WO mol% (e.g., 1-5 mol%, or 1.5-2.5 mol%) PEGylated lipid. In some embodiments, the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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 payload may be at least 70%. 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%). In some embodiments, the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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 payload may be at least 70%. 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). In some embodiments, the lipid-based carrier comprises a TREM (e.g., a TREM described herein) 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 payload may be at least 70%. In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises (i) a TREM (e.g., a TREM described herein); (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 Attorney Docket No.: F2099-7037WO 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. In some embodiments, the lipid-based carrier (or lipid nanoformulation) comprises (i) a TREM (e.g., a TREM described herein); (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 carrier; and (d) a conjugated lipid comprising from 0.5 mol % to 2 mol % of the total lipid present in the lipid- based carrier. 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. 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. 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. The ratio of total lipid components to the payload (e.g., an encapsulated therapeutic agent such as a TREM (e.g., a TREM described herein) can be varied as desired. For example, the total lipid components to the payload (mass or weight) ratio can be from about 10:1 to about 30:1. In some embodiments, the total lipid components to the payload 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 payload 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, Attorney Docket No.: F2099-7037WO 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. Nitrogen:phosphate ratios (N:P ratio) is evaluated at values between 0.1 and 100. The efficiency of encapsulation of a payload such as a protein and/or nucleic acid, describes the amount of protein and/or nucleic acid 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 protein or nucleic acid 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 protein or nucleic acid (e.g., RNA) in a solution. Fluorescence may be used to measure the amount of free protein and/or nucleic acid (e.g., RNA) 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%. Delivery without a carrier A TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition described herein can be administered to a cell without a carrier, e.g., via naked delivery of the TREM, a TREM core fragment or a TREM fragment, a TREM composition or a pharmaceutical TREM composition. In some embodiments, naked delivery as used herein refers to delivery without a carrier. In some embodiments, delivery without a carrier, e.g., naked delivery, comprises delivery with a moiety, e.g., a targeting peptide. In some embodiments, a TREM, a TREM core fragment or a TREM fragment, or TREM composition, or pharmaceutical TREM composition described herein is delivered to a cell without a carrier, e.g., via naked delivery. In some embodiments, the delivery without a carrier, e.g., naked delivery, comprises delivery with a moiety, e.g., a targeting peptide. Attorney Docket No.: F2099-7037WO Use of TREMs A TREM composition (e.g., a pharmaceutical TREM composition described herein) can modulate a function in a cell, tissue or subject. In embodiments, a TREM composition (e.g., a pharmaceutical TREM composition) described herein is contacted with a cell or tissue, or administered to a subject in need thereof, in an amount and for a time sufficient to modulate (increase or decrease) one or more of the following parameters: adaptor function (e.g., cognate or non-cognate adaptor function), e.g., the rate, efficiency, robustness, and/or specificity of initiation or elongation of a polypeptide chain; ribosome binding and/or occupancy; regulatory function (e.g., gene silencing or signaling); cell fate; mRNA stability; protein stability; protein transduction; protein compartmentalization. A parameter may be modulated, e.g., by at least 5% (e.g., at least 10%, 15%, 20%, 25%, 30%, 40%.50%.60%.70%, 80%, 90%, 100%, 150%, 200% or more) compared to a reference tissue, cell or subject (e.g., a healthy, wild-type or control cell, tissue or subject). All references and publications cited herein are hereby incorporated by reference. The following examples are provided to further illustrate some embodiments of the present invention, but are not intended to limit the scope of the invention; it will be understood by their exemplary nature that other procedures, methodologies, or techniques known to those skilled in the art may alternatively be used. ENUMERATED EMBODIMENTS 1. A tRNA effector molecule (TREM) comprising a sequence of Formula (A): [L1]x-[ASt Domain1]-[L2]x-[DH Domain]-[L3]x-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]x-[ASt Domain2]-[L5]x (A), wherein: independently, [L1], [L5], and [VL Domain], are optional; x is 0 or 1; a nucleotide within the TREM sequence comprises a locked nucleic acid (LNA) moiety or 2’,5’-linked nucleotide. Attorney Docket No.: F2099-7037WO 2. The TREM of embodiment 1, wherein the LNA moiety comprises a heterocyclyl ring or cycloalkyl ring between the 2’-position and the 4’-position of the nucleotide sugar within the nucleotide. 3. The TREM of embodiment 2, wherein the heterocyclyl ring comprises an oxygen atom, nitrogen atom, sulfur atom, selenium atom. 4. The TREM of embodiment 2, wherein the heterocyclyl ring or cycloalkyl ring is a five- membered ring, six-membered ring, seven-membered ring, or eight-membered ring. 5. The TREM of any one of embodiments 1-5, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II):
Figure imgf000327_0001
tautomer, or hydrate thereof, wherein: B is a nucleobase; Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and X is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, Attorney Docket No.: F2099-7037WO alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. 6. The TREM of embodiment 5, wherein B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). 7. The TREM of embodiment 5, wherein B is a non-naturally occurring nucleobase. 8. The TREM of any one of embodiments 5-7, wherein B comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). Attorney Docket No.: F2099-7037WO 9. The TREM of embodiment 8, wherein the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. 10. The TREM of embodiment 8, wherein the chemical modification is a nucleotide sugar modification. 11. The TREM of embodiment 8, wherein the chemical modification is a 2’OMe modification. 12. The TREM of embodiment 8, wherein the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification. 13. The TREM of embodiment 8, wherein the chemical modification is a 2’MOE modification. 14. The TREM of embodiment 8, wherein the chemical modification is a 2’-deoxy modification. 15. The TREM of embodiment 8, wherein the chemical modification is present in the internucleotide region (e.g., a backbone modification). 16. The TREM of embodiment 8, wherein the chemical modification is a phosphorothioate modification. 17. The TREM of any one of embodiments 5-7, wherein B does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). 18. The TREM of any one of embodiments 5-17, wherein Y is O. Attorney Docket No.: F2099-7037WO 19. The TREM of any one of embodiments 5-18, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 20. The TREM of any one of embodiments 5-19, wherein R1 is hydrogen. 21. The TREM of any one of embodiments 5-20, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 22. The TREM of any one of embodiments 5-21, wherein R2 is hydrogen. 23. The TREM of any one of embodiments 5-22, wherein R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 24. The TREM of any one of embodiments 5-23, wherein R3 is hydrogen. 25. The TREM of any one of embodiments 5-24, wherein each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 26. The TREM of any one of embodiments 5-25, wherein each of R4a and R4b is independently hydrogen. 27. The TREM of any one of embodiments 5-26, wherein one of L, M, Z, and X is O. 28. The TREM of any one of embodiments 5-27, wherein one of L, M, Z, and X is N(RB)-. 29. The TREM of any one of embodiments 5-28, wherein one of L and X is O. 30. The TREM of any one of embodiments 5-29, wherein one of L and X is N(RB)-. 31. The TREM of any one of embodiments 5-30, wherein X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. Attorney Docket No.: F2099-7037WO 32. The TREM of any one of embodiments 5-31, wherein X is selected from C1-C6 alkylene, O, N(RB), S, and Se. 33. The TREM of any one of embodiments 5-32, wherein X is O. 34. The TREM of any one of the preceding embodiments, wherein TREM comprises a single LNA moiety. 35. The TREM of any one of embodiments 5-33, wherein the TREM comprises a plurality of LNA moieties. 36. The TREM of embodiment 35, wherein the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 LNA moieties. 37. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. 38. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. 39. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L2]-[DH Domain]-[L3]. 40. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L3]-[ACH Domain]. 41. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [VL Domain]. Attorney Docket No.: F2099-7037WO 42. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [TH Domain]. 43. The TREM of any one of the preceding embodiments, wherein the LNA moiety is present in the [L4]-[ASt Domain2]-[L5]. 44. The TREM of any one of the preceding embodiments, wherein the TREM has a sequence selected from a sequence provided in FIG.1. 45. The TREM of any one of the preceding embodiments, wherein the TREM comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in FIG.1. 46. The TREM of any one of the preceding embodiments, wherein the TREM comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from a TREM provided in FIG.1. 47. The TREM of any one of the preceding embodiments, wherein the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, LNA, or phosphorothiorate modifications). 48. The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of any one of SEQ ID NOs: 626-793. 49. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-a): Attorney Docket No.: F2099-7037WO or hydrate thereof, wherein:
Figure imgf000333_0001
, or ; each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; Attorney Docket No.: F2099-7037WO each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. 50. The TREM of embodiment 49, wherein Y is selected from O or N(RB). 51. The TREM of any one of embodiments 49-50, wherein Y is O. 52. The TREM of any one of embodiments 49-51, wherein R1 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 53. The TREM of any one of embodiments 49-52, wherein R1 is hydrogen. 54. The TREM of any one of embodiments 49-53, wherein R2 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 55. The TREM of any one of embodiments 49-54, wherein R2 is hydrogen. 56. The TREM of any one of embodiments 49-55, wherein R3 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 57. The TREM of any one of embodiments 49-56, wherein R3 is hydrogen. Attorney Docket No.: F2099-7037WO 58. The TREM of any one of embodiments 49-57, wherein each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 59. The TREM of any one of embodiments 49-58, wherein each of R4a and R4b is independently hydrogen. 60. The TREM of any one of embodiments 49-59, wherein one of L, M, Z, and X is O. 61. The TREM of any one of embodiments 49-60, wherein one of L, M, Z, and X is N(RB)-. 62. The TREM of any one of embodiments 49-61, wherein one of L and X is O. 63. The TREM of any one of embodiments 49-62, wherein one of L and X is N(RB)-. 64. The TREM of any one of embodiments 49-63, wherein X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. 65. The TREM of any one of embodiments 49-64, wherein X is selected from C1-C6 alkylene, O, N(RB), -S-, and -Se-. 66. The TREM of any one of embodiments 49-65, wherein X is O. 67. The TREM of any one of embodiments 49-66, wherein each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 68. The TREM of any one of embodiments 49-67, wherein each of Ria and Rib is independently hydrogen. 69. The TREM of any one of embodiments 49-68, wherein each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or halo. Attorney Docket No.: F2099-7037WO 70. The TREM of any one of embodiments 49-69, wherein each of Rii and Riii is independently hydrogen. 71. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-b): or hydrate thereof, wherein:
Figure imgf000336_0001
Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; Attorney Docket No.: F2099-7037WO each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. 72. The TREM of embodiment 71, wherein Y is selected from O or N(RB). 73. The TREM of any one of embodiments 71-72, wherein Y is O. 74. The TREM of any one of embodiments 71-73, wherein R1 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 75. The TREM of any one of embodiments 71-74, wherein R1 is hydrogen. 76. The TREM of any one of embodiments 71-75, wherein R2 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. Attorney Docket No.: F2099-7037WO 77. The TREM of any one of embodiments 71-76, wherein R2 is hydrogen. 78. The TREM of any one of embodiments 71-77, wherein R3 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 79. The TREM of any one of embodiments 71-78, wherein R3 is hydrogen. 80. The TREM of any one of embodiments 71-79, wherein each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 81. The TREM of any one of embodiments 71-80, wherein each of R4a and R4b is independently hydrogen. 82. The TREM of any one of embodiments 71-81, wherein one of L, M, Z, and X is O. 83. The TREM of any one of embodiments 71-82, wherein one of L, M, Z, and X is N(RB)-. 84. The TREM of any one of embodiments 71-83, wherein one of L and X is O. 85. The TREM of any one of embodiments 71-84, wherein one of L and X is N(RB)-. 86. The TREM of any one of embodiments 71-85, wherein X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. 87. The TREM of any one of embodiments 71-86, wherein X is selected from C1-C6 alkylene, -O-, -N(RB)-, -S-, and -Se-. 88. The TREM of any one of embodiments 71-87, wherein X is O. Attorney Docket No.: F2099-7037WO 89. The TREM of any one of embodiments 71-88, wherein each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 90. The TREM of any one of embodiments 71-89, wherein each of Ria and Rib is independently hydrogen. 91. The TREM of any one of embodiments 71-90, wherein each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or halo. 92. The TREM of any one of embodiments 71-91, wherein each of Rii and Riii is independently hydrogen. 93. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-c):
Figure imgf000339_0001
tautomer, or hydrate thereof, wherein: Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; Attorney Docket No.: F2099-7037WO each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. 94. The TREM of embodiment 93, wherein Y is selected from O or N(RB). 95. The TREM of any one of embodiments 93-94, wherein Y is O. Attorney Docket No.: F2099-7037WO 96. The TREM of any one of embodiments 93-95, wherein R1 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 97. The TREM of any one of embodiments 93-96, wherein R1 is hydrogen. 98. The TREM of any one of embodiments 93-97, wherein R2 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 99. The TREM of any one of embodiments 93-98, wherein R2 is hydrogen. 100. The TREM of any one of embodiments 93-99, wherein R3 is hydrogen, C1-C6 alkyl, C1- C6 heteroalkyl, C1-C6 haloalkyl, or halo. 101. The TREM of any one of embodiments 93-100, wherein R3 is hydrogen. 102. The TREM of any one of embodiments 93-101, wherein each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 103. The TREM of any one of embodiments 93-102, wherein each of R4a and R4b is independently hydrogen. 104. The TREM of any one of embodiments 93-103, wherein one of L, M, Z, and X is O. 105. The TREM of any one of embodiments 93-104, wherein one of L, M, Z, and X is N(RB)-. 106. The TREM of any one of embodiments 93-105, wherein one of L and X is O. 107. The TREM of any one of embodiments 93-106, wherein one of L and X is N(RB)-. Attorney Docket No.: F2099-7037WO 108. The TREM of any one of embodiments 93-107, wherein X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. 109. The TREM of any one of embodiments 93-108, wherein X is selected from C1-C6 alkylene, -O-, -N(RB)-, -S-, and -Se-. 110. The TREM of any one of embodiments 93-109, wherein X is O. 111. The TREM of any one of embodiments 93-110, wherein each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 112. The TREM of any one of embodiments 93-111, wherein each of Ria and Rib is independently hydrogen. 113. The TREM of any one of embodiments 93-112, wherein each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or halo. 114. The TREM of any one of embodiments 93-113, wherein each of Rii and Riii is independently hydrogen. 115. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-d):
Figure imgf000342_0001
tautomer, or hydrate thereof, wherein: Y is C(R’), O, or N(RB); Attorney Docket No.: F2099-7037WO each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and Z is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; Ri is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; Attorney Docket No.: F2099-7037WO provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. 116. The TREM of embodiment 115, wherein Y is selected from O or N(RB). 117. The TREM of any one of embodiments 115-116, wherein Y is O. 118. The TREM of any one of embodiments 115-117, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 119. The TREM of any one of embodiments 115-118, wherein R1 is hydrogen. 120. The TREM of any one of embodiments 115-119, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 121. The TREM of any one of embodiments 115-120, wherein R2 is hydrogen. 122. The TREM of any one of embodiments 115-121, wherein R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 123. The TREM of any one of embodiments 115-122, wherein R3 is hydrogen. 124. The TREM of any one of embodiments 115-123, wherein each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 125. The TREM of any one of embodiments 115-124, wherein each of R4a and R4b is independently hydrogen. 126. The TREM of any one of embodiments 115-125, wherein one of L, M, Z, and X is O. Attorney Docket No.: F2099-7037WO 127. The TREM of any one of embodiments 115-126, wherein one of L, M, Z, and X is N(RB)-. 128. The TREM of any one of embodiments 115-127, wherein one of L and X is O. 129. The TREM of any one of embodiments 115-128, wherein one of L and X is N(RB)-. 130. The TREM of any one of embodiments 5115-129, wherein X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. 131. The TREM of any one of embodiments 115-130, wherein X is selected from C1-C6 alkylene, -O-, -N(RB)-, -S-, and -Se-. 132. The TREM of any one of embodiments 115-131, wherein X is O. 133. The TREM of any one of embodiments 115-132, wherein Ri is selected from hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 134. The TREM of any one of embodiments 115-133, wherein Ri is hydrogen. 135. The TREM of any one of embodiments 115-134, wherein each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or halo. 136. The TREM of any one of embodiments 115-135, wherein each of Rii and Riii is independently hydrogen. 137. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-e): Attorney Docket No.: F2099-7037WO or hydrate thereof, wherein:
Figure imgf000346_0001
a Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; Attorney Docket No.: F2099-7037WO each of x and y is independently 0, 1, or 2; n is 1, 2, 3, 4, 5, or 6; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 138. The TREM of embodiment 137, wherein Y is O. 139. The TREM of any one of embodiments 137-138, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 140. The TREM of any one of embodiments 137-139, wherein R1 is hydrogen. 141. The TREM of any one of embodiments 137-140, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 142. The TREM of any one of embodiments 137-141, wherein R2 is hydrogen. 143. The TREM of any one of embodiments 137-142, wherein R3 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 144. The TREM of any one of embodiments 137-143, wherein R3 is hydrogen. 145. The TREM of any one of embodiments 137-144, wherein each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 146. The TREM of any one of embodiments 137-145, wherein each of R4a and R4b is independently hydrogen. 147. The TREM of any one of embodiments 137-146, wherein each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. Attorney Docket No.: F2099-7037WO 148. The TREM of any one of embodiments 137-147, wherein each of R5a and R5b is independently hydrogen. 149. The TREM of any one of embodiments 137-148, wherein one of R5a and R5b is independently methyl. 150. The TREM of any one of embodiments 137-149, wherein X is selected from C1-C6 alkylene, O, N(RB), S(RD)x-, and -Se(RE)y-. 151. The TREM of any one of embodiments 137-150, wherein X is selected from C1-C6 alkylene, O, N(RB), S, and Se. 152. The TREM of any one of embodiments 137-151, wherein X is O. 153. The TREM of any one of embodiments 137-152, wherein X is N(RB). 154. The TREM of any one of the preceding embodiments, wherein n is selected from 1, 2, or 3. 155. The TREM of any one of the preceding embodiments, wherein n is 1. 156. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-e-i) or (II-e-ii): e-ii)
Figure imgf000348_0001
Attorney Docket No.: F2099-7037WO 157. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-f): or hydrate thereof, wherein:
Figure imgf000349_0001
a each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; Attorney Docket No.: F2099-7037WO RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; n is 1, 2, 3, 4, 5, or 6; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 158. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-f-i) or (II-f-ii): ii)
Figure imgf000350_0001
159. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety having a structure of Formula (II-f) is selected from: i),
Figure imgf000350_0002
Attorney Docket No.: F2099-7037WO 160. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-g): or hydrate thereof, wherein:
Figure imgf000351_0001
a each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; n is 1, 2, 3, 4, 5, or 6; and Attorney Docket No.: F2099-7037WO “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 161. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-g-i) or Formula (II- g-ii):
Figure imgf000352_0001
162. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-g-iii): iii)
Figure imgf000352_0002
or hydrate thereof. 163. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-l):
Attorney Docket No.: F2099-7037WO Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; M is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. Attorney Docket No.: F2099-7037WO 164. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-m): , or hydrate thereof, wherein:
Figure imgf000354_0001
a each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R6a and R6b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; Attorney Docket No.: F2099-7037WO RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 165. The TREM of one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety having a structure of Formula (II-l) is selected from:
Figure imgf000355_0001
166. The TREM of one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-m-i):
Figure imgf000355_0002
tautomer, or hydrate thereof. 167. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-r): Attorney Docket No.: F2099-7037WO or hydrate thereof, wherein:
Figure imgf000356_0001
a Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R6a and R6b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R7a and R7b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or Attorney Docket No.: F2099-7037WO RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the the TREM. 168. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequencing comprising the LNA moiety having a structure of Formula (II-r), is selected from Formula (II-s) or Formula (II-t): t),
Figure imgf000357_0001
169. The TREM of embodiment 1, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (II):
Figure imgf000357_0002
tautomer, or hydrate thereof, wherein: B is a nucleobase; Y is C(R’), O, or N(RB); Attorney Docket No.: F2099-7037WO each of R1, R2, and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the the TREM. 170. The TREM of embodiment 169, wherein B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). 171. The TREM of any one of embodiments 169-170, wherein B is a non-naturally occurring nucleobase. 172. The TREM of any one of embodiments 169-171, wherein the compound of Formula (III) further comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). Attorney Docket No.: F2099-7037WO 173. The TREM of embodiment 172, wherein the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. 174. The TREM of any one of embodiments 169-173, wherein the chemical modification is a nucleotide sugar modification. 175. The TREM of embodiment 174, wherein the chemical modification is a 2’OMe modification. 176. The TREM of embodiment 174, wherein the chemical modification is a 2’halo (e.g., 2’F or 2’Cl) modification. 176. The TREM of embodiment 174, wherein the chemical modification is a 2’MOE modification. 177. The TREM of embodiment 174, wherein the chemical modification is a 2’-deoxy modification. 178. The TREM of embodiment 173, wherein the chemical modification is present in the internucleotide region (e.g., a backbone modification). 179. The TREM of embodiment 173, wherein the chemical modification is a phosphorothioate modification. 180. The TREM of any one of embodiments 169-179, wherein the compound of Formula (III) does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). 181. The TREM of any one of embodiments 169-180, wherein Y is O. Attorney Docket No.: F2099-7037WO 182. The TREM of any one of embodiments 169-181, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 183. The TREM of any one of embodiments 169-182, wherein R1 is hydrogen. 184. The TREM of any one of embodiments 169-183, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 185. The TREM of any one of embodiments 169-184, wherein R2 is hydrogen. 186. The TREM of any one of embodiments 169-185, wherein R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 187. The TREM of any one of embodiments 169-186, wherein R4 is hydrogen. 188. The TREM of any one of embodiments 169-187, wherein each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 189. The TREM of any one of embodiments 169-188, wherein each of R3a and R3b is independently hydrogen. 190. The TREM of any one of the preceding embodiments, wherein TREM comprises a single 2’,5’-linked nucleotide moiety. 191. The TREM of any one of embodiments 169-189, wherein the TREM comprises a plurality of 2’,5’-linked nucleotide moieties. 192. The TREM of embodiment 191, wherein the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 252’,5’-linked nucleotides. Attorney Docket No.: F2099-7037WO 193. The TREM of any one of the preceding embodiments, wherein the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]-[L2]. 194. The TREM of any one of the preceding embodiments, wherein the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]-[L2]. 195. The TREM of any one of the preceding embodiments, wherein the 2’,5’-linked nucleotide is present in the [L2]-[DH Domain]-[L3]. 196. The TREM of any one of the preceding embodiments, wherein the 2’,5’-linked nucleotide is present in the [L3]-[ACH Domain]. 197. The TREM of any one of the preceding embodiments, wherein the 2’,5’-linked nucleotide is present in the [VL Domain]. 198. The TREM of any one of the preceding embodiments, wherein the 2’,5’-linked nucleotide is present in the [TH Domain]. 199. The TREM of any one of the preceding embodiments, wherein the 2’,5’-linked nucleotide is present in the [L4]-[ASt Domain2]-[L5]. 200. The TREM of any one of the preceding embodiments, wherein the TREM has a sequence selected from a sequence provided in FIG.1. 201. The TREM of any one of the preceding embodiments, wherein the TREM comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in FIG.1. 202. The TREM of any one of the preceding embodiments, wherein the TREM comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from a TREM provided in FIG.1. Attorney Docket No.: F2099-7037WO 203. The TREM of any one of the preceding embodiments, wherein the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, or phosphorothiorate modifications). 204. The TREM of any one of the preceding embodiments, wherein the TREM comprises the nucleotide sequence of 205. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-a):
Figure imgf000362_0001
or hydrate thereof, wherein: B is a nucleobase; Y is C(R’), O, or N(RB); each of R1, R2, and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R3a is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R3c is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; Attorney Docket No.: F2099-7037WO R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 206. The TREM of embodiment 205, wherein Y is O. 207. The TREM of any one of embodiments 205-206, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 208. The TREM of any one of embodiments 205-207, wherein R1 is hydrogen. 209. The TREM of any one of embodiments 205-208, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 210. The TREM of any one of embodiments 205-209, wherein R2 is hydrogen. 211. The TREM of any one of embodiments 205-210, wherein R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 212. The TREM of any one of embodiments 205-211, wherein R4 is hydrogen. Attorney Docket No.: F2099-7037WO 213. The TREM of any one of embodiments 205-212, wherein R3a is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 214. The TREM of any one of embodiments 205-213, wherein R3a is independently hydrogen. 215. The TREM of any one of embodiments 205-214, wherein each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 216. The TREM of any one of embodiments 205-215, wherein each of R5a and R5b is independently hydrogen. 217. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-a-i) or Formula (III-a-ii): a-ii)
Figure imgf000364_0001
218. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-b):
Attorney Docket No.: F2099-7037WO C2-C6 alkenyl, C2-C6
Figure imgf000365_0001
alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, - or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, or C1-C6 haloalkyl; Attorney Docket No.: F2099-7037WO each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 219. The TREM of embodiment 218, wherein Y is O. 220. The TREM of any one of embodiments 218-219, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 221. The TREM of any one of embodiments 218-220, wherein R1 is hydrogen. 222. The TREM of any one of embodiments 218-221, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 223. The TREM of any one of embodiments 218-222, wherein R2 is hydrogen. 224. The TREM of any one of embodiments 218-223, wherein R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 225. The TREM of any one of embodiments 218-224, wherein R4 is hydrogen. 226. The TREM of any one of embodiments 218-225, wherein each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 227. The TREM of any one of embodiments 218-226, wherein each of R3a and R3b is independently hydrogen. 228. The TREM of any one of embodiments 218-227, wherein each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. Attorney Docket No.: F2099-7037WO 229. The TREM of any one of embodiments 218-228, wherein each of R5a and R5b is independently hydrogen. 230. The TREM of any one of embodiments 218-229, wherein each of Ria and Rib is independently hydrogen, C1-C6 alkyl or C1-C6-heteroalkyl. 231. The TREM of any one of embodiments 218-230, wherein each of Ria and Rib is independently hydrogen. 232. The TREM of any one of embodiments 218-231, wherein each of Rii and Riii is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. 233. The TREM of any one of embodiments 218-232, wherein each of Rii and Riii is independently hydrogen. 234. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-c):
Figure imgf000367_0001
thereof, wherein: Y is C(R’), O, or N(RB); each of R1, R2, and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; Attorney Docket No.: F2099-7037WO each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, or C1-C6 haloalkyl; Rii is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl; Riii is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM 235. The TREM of embodiment 234, wherein Y is O. 236. The TREM of any one of embodiments 234-235, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 237. The TREM of any one of embodiments 234-236, wherein R1 is hydrogen. Attorney Docket No.: F2099-7037WO 238. The TREM of any one of embodiments 234-237, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 239. The TREM of any one of embodiments 234-238, wherein R2 is hydrogen. 240. The TREM of any one of embodiments 234-239, wherein R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 241. The TREM of any one of embodiments 234-240, wherein R4 is hydrogen. 242. The TREM of any one of embodiments 234-241, wherein each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 243. The TREM of any one of embodiments 234-242, wherein each of R3a and R3b is independently hydrogen. 244. The TREM of any one of embodiments 234-243, wherein each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 245. The TREM of any one of embodiments 234-244, wherein each of R5a and R5b is independently hydrogen. 246. The TREM of any one of embodiments 234-245, wherein each of Ria and Rib is independently hydrogen, C1-C6 alkyl or C1-C6-heteroalkyl. 247. The TREM of any one of embodiments 234-246, wherein each of Ria and Rib is independently hydrogen. 248. The TREM of any one of embodiments 234-247, wherein Rii is hydrogen or C1-C6 alkyl. 249. The TREM of any one of embodiments 234-248, wherein Rii is hydrogen. Attorney Docket No.: F2099-7037WO 250. The TREM of any one of embodiments 234-249, wherein Riii is hydrogen or C1-C6 alkyl. 251. The TREM of any one of embodiments 234-250, wherein Riii is hydrogen. 252. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-d):
Figure imgf000370_0001
or hydrate thereof, wherein: Y is C(R’), O, or N(RB); each of R1, R2, and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or Attorney Docket No.: F2099-7037WO RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of Ria and Rib is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 253. The TREM of embodiment 252, wherein Y is O. 254. The TREM of any one of embodiments 252-253, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 255. The TREM of any one of embodiments 252-254, wherein R1 is hydrogen. 256. The TREM of any one of embodiments 252-255, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 257. The TREM of any one of embodiments 252-256, wherein R2 is hydrogen. 258. The TREM of any one of embodiments 252-257, wherein R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 259. The TREM of any one of embodiments 252-258, wherein R4 is hydrogen. 260. The TREM of any one of embodiments 252-259, wherein each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. Attorney Docket No.: F2099-7037WO 261. The TREM of any one of embodiments 252-260, wherein each of R3a and R3b is independently hydrogen. 262. The TREM of any one of embodiments 252-261, wherein each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 263. The TREM of any one of embodiments 252-262, wherein each of R5a and R5b is independently hydrogen. 264. The TREM of any one of embodiments 252-263, wherein each of Ria and Rib is independently hydrogen, C1-C6 alkyl or C1-C6-heteroalkyl. 265. The TREM of any one of embodiments 252-264, wherein each of Ria and Rib is independently hydrogen. 266. The TREM of any one of embodiments 252-265, wherein each of Rii and Riii is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. 267. The TREM of any one of embodiments 252-266, wherein each of Rii and Riii is independently hydrogen. 268. The TREM of any one of the preceding embodiments, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-e):
Attorney Docket No.: F2099-7037WO or a salt, stereoisomer, tautomer, or hydrate thereof, wherein: Y is C(R’), O, or N(RB); each of R1, R2, and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; Ri is hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, or C1-C6 haloalkyl; each of Rii and Riii is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 269. The TREM of embodiment 268, wherein Y is O. 270. The TREM of any one of embodiments 268-269, wherein R1 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. Attorney Docket No.: F2099-7037WO 271. The TREM of any one of embodiments 268-270, wherein R1 is hydrogen. 272. The TREM of any one of embodiments 268-271, wherein R2 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 273. The TREM of any one of embodiments 268-272, wherein R2 is hydrogen. 274. The TREM of any one of embodiments 268-273, wherein R4 is hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 275. The TREM of any one of embodiments 268-274, wherein R4 is hydrogen. 276. The TREM of any one of embodiments 268-275, wherein each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 277. The TREM of any one of embodiments 268-276, wherein each of R3a and R3b is independently hydrogen. 278. The TREM of any one of embodiments 268-277, wherein each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, or halo. 279. The TREM of any one of embodiments 268-278, wherein each of R5a and R5b is independently hydrogen. 280. The TREM of any one of embodiments 268-279, wherein Ri is hydrogen, C1-C6 alkyl or C1-C6-heteroalkyl. 281. The TREM of any one of embodiments 268-280, wherein Ri is hydrogen. Attorney Docket No.: F2099-7037WO 282. The TREM of any one of embodiments 268-281, wherein each of Rii and Riii is independently hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl. 283. The TREM of any one of embodiments 268-282, wherein each of Rii and Riii is independently hydrogen. 284. A pharmaceutical composition comprising a TREM of any one of the preceding embodiments. 285. The pharmaceutical composition of embodiment 284, further comprising a pharmaceutically acceptable component, e.g., an excipient. 286. A lipid nanoparticle formulation comprising a TREM of any one of embodiments 1-283. 287. A lipid nanoparticle formulation comprising a pharmaceutical composition of embodiment 284. 288. A method of treating a subject having a disease or disorder associated with a PTC comprising administering to the subject a TREM, TREM core fragment, or TREM fragment described herein (e.g., a TREM of any one of embodiments 1-283), thereby treating the subject having the disease or disorder. 289. The method of embodiment 288, wherein the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease. EXAMPLES Table of Contents for Examples Example 1 Synthesis of modified TREMs
Figure imgf000375_0001
Attorney Docket No.: F2099-7037WO Example 5 Deprotection of synthesized TREM Example 6 Characterization of chemically modified TREMs for readthrough of a
Figure imgf000376_0001
Generally, TREM molecules (e.g., modified TREMs) comprising a locked nucleic acid (LNA) moiety or a 2’,5’-linked nucleotide described herein may be chemically synthesized and purified by HPLC according to standard solid phase synthesis methods using phosphoramidite chemistry (see, e.g., Scaringe S. et al. (2004) Curr Protoc Nucleic Acid Chem, 2.10.1-2.10.16; Usman N. et al. (1987) J. Am. Chem. Soc, 109, 7845-7854). The TREMs may also contain additional non-naturally occurring modifications, such as a 2’-methoxy (2’OMe), 2’fluoro (2’F), 2’-methoxyethyl (2’-MOE), or phosphorothioate (PS) modification. For clarity, the arginine non- cognate TREM molecule named TREM-Arg-TGA contains the sequence of ARG-UCU-TREM body but with the anticodon sequence corresponding to UCA instead of UCU (i.e., SEQ ID NO: 622). To make the LNA-containing TREMs, the following exemplary phosphoramidites were used: 5’-dimethoxytrityl-5-methyl-(2’-O, 4’-C methylene)-uridine,3’-[(2-cyanoethyl)-(N,N- diisopropyl)]-phosphoramidite, 5’-dimethoxytrityl-N-benzoyl-(2’-O, 4’-C methylene)- adenosine,3’-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 5’-dimethoxytrityl-N- dimethylformamidine-(2’-O, 4’-C methylene)-guanosine, 3’-[(2-cyanoethyl)-(N,N-diisopropyl)]- phosphoramidite, and 5’-dimethoxytrityl-N-benzoyl-5-methyl-(2’-O, 4’-C methylene)- cytidine,3’-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite. To make the 2’,5’-linked nucleotide-containing TREMs, the following exemplary phosphoramidites were used: 5'-O-DMT-3'-TBDMS-N6-Bz-Adenosine 2'-CE phosphoramidite, 5'-O-DMT-3'-TBDMS-N4-Acetyl-Cytidine 2'-CE phosphoramidite, 5'-O-DMT-3'-O-TBDMS- N2-iBu-Guanosine 2'-CE phosphoramidite, and 5'-O-DMT-3'-TBDMS-Uridine 2'-CE phosphoramidite. To make the 2’OMe modified TREMs, the following 2’-O-methyl phosphoramidites were used: (5’-O-dimethoxytrityl-N6-(benzoyl)-2’-O-methyl-adenosine-3’-O-(2-cyanoethyl- Attorney Docket No.: F2099-7037WO N,N-diisopropylamino) phosphoramidite, 5’-O-dimethoxy-trityl-N4-(acetyl)-2’-O-methyl- cytidine-3’-O-(2-cyanoethyl-N,N-diisopropyl-amino) phosphoramidite, (5’- O -dimethoxytrityl- N2-(isobutyryl)-2’-O-methyl-guanosine-3’-O-(2-cyano-ethyl-N,N-diisopropylamino)- phosphoramidite, and 5’-O-dimethoxy-trityl-2’-O-methyluridine-3’- O -(2-cyanoethyl-N,N- diisopropylamino)phosphoramidite. To make the 2’-deoxy and 2’-F modified TREMs, analogous 2’-deoxy and 2’-fluoro-phosphoramidites with the same protecting groups as the 2’-O-methyl RNA amidites were used. To make the 2’-MOE modified TREMs, the following 2’-MOE- phosphoramidites were used: 5’-O-(4,4’-Dimethoxytrityl)-2’-O-methoxyethyl-N6-benzoyl- adenosine -3’-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 5’-O-(4,4’- Dimethoxytrityl)-2’-O-methoxyethyl-5-methyl-N4-benzoyl- cytidine-3’-O-[(2-cyanoethyl)- (N,N-diisopropyl)]-phosphoramidite, 5’-O-(4,4’-Dimethoxytrityl)-2’-O-methoxyethyl-N2- isobutyryl- guanosine-3’-O-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, 5’-O-(4,4’- Dimethoxytrityl)-2’-O-methoxyethyl-5-methyl-uridine-3’-O-[(2-cyanoethyl)-(N,N-diisopropyl)]- phosphoramidite. During the oligonucleotide synthesis via this phosphoramidites approach, the phosphorothioate was introduced by oxidizing the phosphite triester using a sulfur transfer reagent, such as tetraethylthiuram disulfide (TETD), bis(O,O-diisopropoxy phosphinothioyl) disulfide (Stec’s reagent), 3H-1,2-benzodithiol-3-one-1,1,-dioxide (Beaucage reagent), phenylacetyl disulfide (PADS), 3-ethoxy-1,2,4-dithiazoline-5-one (EDITH), 1,2-dithiazole-5- thione (xanthane hydride or ADTT), 3-((dimethylamino-methylidene)amino)-3H-1,2,4- dithiazole-3-thione (DDTT), dimethylthiuram disulfide (DTD), 3-phenyl-1,2,4-dithiazoline-5- one (PolyOrg Sulfa or POS). FIG.1 describes a series of singly and multiply modified TREMs synthesized according to this procedure. The sequences of each of these TREMs are provided in the table, wherein r: ribonucleotide; LNA: locked nucleic acid; [25r] or 2_5_r: 2’,5’-linked nucleotide; m: 2’-OMe; *: PS linkage; f: 2’-fluoro; moe: 2’-moe; d: deoxyribonucleotide; [m5C] or 5MeC: 5- methylcytosine. Thus, for example, mA represents 2’-O-methyl adenosine, moe5MeC represents 2’-MOE nucleotide with 5-methylcytosine nucleobase, and dA represents an adenosine deoxyribonucleotide. Example 2: HPLC and MS analysis of modified TREMs Attorney Docket No.: F2099-7037WO Chemically modified TREM molecules may be analyzed by HPLC, for example, to evaluate the purity and homogeneity of the compositions. A Waters Aquity UPLC system using a Waters BEH C18 column (2.1 mm x 50 mm x 1.7 μm) may be used for this analysis. Samples may be prepared by dissolving 0.5 nmol of the TREM in 75 μL of water and injecting 2 μL of the solution. The buffers used may be 50 mM dimethylhexylammonium acetate with 10% CH3CN (acetonitrile) as buffer A and 50 mM dimethylhexylammonium acetate with 75% CH3CN as buffer B (gradient 25-75% buffer B over 5 mins), with a flow rate of 0.5 mL/min at 60 °C. ESI-LCMS data for the chemically modified TREMs may be acquired on a Thermo Ultimate 3000-LTQ-XL mass spectrometer. FIG.1 describe a series of singly and multiply modified TREMs synthesized according to the protocol outlined in Example 1. The calculated and detected molecular weights for each sequence were determined as outlined herein. Example 3: Analysis of modified TREMs via anion-exchange HPLC This example describes the quality control of a synthesized TREM via anion-exchange HPLC. Using the Dionex DNA-Pac–PA-100 column, a gradient is employed using HPLC buffer A and HPLC buffer B.0.5 ODUs of a sample that has been dissolved in H2O or Tris buffer, pH 7.5 is injected onto the gradient. The gradient employed is based on oligonucleotide length and can be applied according to Table 13. The parameters provided in Table 14 can be used to program a linear gradient on the HPLC analyzer. Table 13: Oligonucleotide length and gradient percentages Length Gradient
Figure imgf000378_0001
Table 14: Parameters for a linear gradient on HPLC analyzer Attorney Docket No.: F2099-7037WO Time Flow % Buffer % Buffer (min) (mL/min) A B
Figure imgf000379_0001
Example 4: Analysis of TREMs via PAGE Purification and Analysis This example describes the quality control of a synthesized TREM via PAGE Purification and Analysis. Gel purification and analysis of 2’-ACE protected RNA follows standard protocols for denaturing PAGE (Ellington and Pollard (1998) In Current Protocols in Molecular Biology, Chanda, V). Briefly, the 2’-ACE protected oligo is resuspended in 200 mL of gel loading buffer. Invitrogen™ NuPAGE™ 4-12% Bis-Tris Gels or similar gel is prepared in gel apparatus. Samples are loaded and gel ran at 50-120 W, maintaining the apparatus at 40°C. When complete, the gel is exposed to ultraviolet (UV) light at 254 nm to visualize the purity of the RNA using UV shadowing. If necessary, the desired gel band is excised with a clean razor blade. The gel slice is crushed and 0.3M NaOAc elution buffer is added to the gel particles, and soaked overnight. The mixture is decanted and filtered through a Sephadex column such as Nap-10 or Nap-25. Example 5: Deprotection of synthesized TREM This example describes the deprotection of a TREM made according to an in vitro synthesis method. The 2’-protecting groups are removed using 100 mM acetic acid, pH 3.8. The formic acid and ethylene glycol byproducts are removed by incubating at 60°C for 30 min Attorney Docket No.: F2099-7037WO followed by lyophilization or SpeedVac-ing to dryness. After this final deprotection step, the oligonucleotides are ready for use. Example 6. Characterization of chemically modified TREMs for readthrough of a premature termination codon (PTC) in a reporter protein This example describes an assay to test the ability of a non-cognate chemically modified TREM to readthrough a PTC in a cell line expressing a reporter protein having a PTC. This protocol describes analysis of chemically modified arginine and glutamate non-cognate TREM (i.e., Arg-TGA, Glu-TAG), though a non-cognate TREM specifying any one of the other amino acids can also be used. A cell line engineered to stably express the NanoLuc reporter construct containing a premature termination codon (PTC) may be generated using the FlpIn and/or JumpIN cell system (ThermoFisher Scientific, USA) according to the manufacturer’s instructions. The NanoLuc reporter can also be modified with a destabilizing PEST domain at its C-terminus to reduce the protein half-life. Delivery of the chemically modified TREMs into the NanoLuc reporter cells is carried out via a reverse transfection reaction using lipofectamine RNAiMAX (ThermoFisher Scientific, USA) according to manufacturer instructions. Briefly, 0.15uL of a 5uM solution of chemically modified TREM sample are diluted in a 7.35uL RNAiMAX/OptiMEM mixture. After 30min gentle mixing at room temperature, the 7.5uL TREM/transfection mixture is added to a 384-well plate and kept still for 20-30min before adding the cells stably expressing the NanoLuc reporter construct containing a PTC. The NanoLuc reporter cells are harvested and diluted to 4× 105 cells/mL in complete growth medium, and 30uL of the diluted cell suspension is added and mixed to the plate containing the TREM. To monitor the efficacy of the chemically modified TREM to read through the PTC in the reporter construct at 24 or 48 hours after TREM delivery into cells, a NanoGlo bioluminescent assay (Promega, USA) may be performed according to manufacturer instruction. Briefly, for cells that are to be assayed after 48 hours, 30uL complete growth medium is added to the 384- well plate 24 hours post-transfection to dilute the transfection reagent for cell health. At the time of harvest, either 24- or 48-hours post-transfection, cell media is removed and plates are stored at -80C. To perform the assay, thaw plates are thawed on ice for 1 hour, then transfered to room temperature and allowed to equilibrate. Add 15 uL room-temperature complete growth media to each well. NanoGlo reagent is prepared by mixing the buffer with substrate in a 50:1 ratio.15uL Attorney Docket No.: F2099-7037WO of mixed NanoGlo reagent is added to each well of the 384-well plate and mixed on the shaker at 600rpm for 10min. Then the plate is centrifuged at 100g for 1min, followed by a 5min incubation step at room temperature before measuring sample bioluminescence. As a positive control, a host cell expressing the NanoLuc reporter construct with a PTC is used and transfected with a previously-identified unmodified, active TREM. As a negative control, a host cell expressing the NanoLuc reporter construct with a PTC is used, but no TREM is transfected. The efficacy of the chemically modified TREMs is measured as a ratio of the NanoLuc luminescence in the experimental sample to the NanoLuc luminescence of the positive control or as a ratio of the NanoLuc luminescence in the experimental sample to the NanoLuc luminescence of the negative control. It is expected that if the sample TREM is functional, it may be able to read-through the stop mutation in the NanoLuc reporter and produce a luminescent reading higher than the luminescent reading measured in the negative control. If the sample TREM is not functional, the stop mutation is not rescued, and luminescence less or equal to the negative control is detected. The impacts of chemical modifications were evaluated in singly and multiply modified TREM sequences and are summarized in FIG.1. In this figure, the TREMs are annotated as follows: r: ribonucleotide; m: 2’-OMe; *: PS linkage; f: 2’-fluoro; moe: 2’-moe; d: deoxyribonucleotide; 5MeC: 5-methylcytosine. Thus, for example, mA represents 2’-O-methyl adenosine, moe5MeC represents 2’-MOE nucleotide with 5-methylcytosine nucleobase, and dA represents an adenosine deoxyribonucleotide. FIG.1 also summarizes the results of the activity screen in column “A” for measurements made using NanoLuc reporter cells at 48 hours post-transfection, which reported as log2 fold changes compared with the appropriate unmodified TREM, wherein “1” indicates less than a 1 log2 fold change; “2” indicates greater than or equal to 1 and less than 3.32 log2 fold change; and “3” indicates greater than or equal to 3.32 log2 fold change. The results show that certain modifications were tolerated at many positions, but particular sites were sensitive to modification or exhibited improved activity when modified Example 7: Characterization of chemically modified TREMs for readthrough of a premature termination codon (PTC) in a disease reporter cell line This example describes an assay to test the ability of an exemplary TREM to readthrough a PTC in a cell line expressing a disease reporter protein bearing the PTC. Host cell modification Attorney Docket No.: F2099-7037WO A cell line engineered to stably express a HaloTag and HiBiT-tagged disease reporter construct containing a premature termination codon (PTC), such as Factor IX at position 248 (FIXR248X), Factor IX at position 29 (FIXR29X), Factor IX at position 333 (FIXR333X), or Rhodopsin at position 334 (RhoS334X) was generated using the Jump-In system according to manufacturer’s instructions. Briefly, Jump-In GripTite HEK293 (Thermo Scientific A14150) cells were co- transfected with an expression vector containing the disease reporter, such as pJTI-R4-DEST- CMV-FIX-R298X-HaloTag-HiBiT-pA for FIXR298X to make the Factor IX disease reporter expressing cell line, and a pJTI-R4-Int PhiC31 integrase expression vector using Lipofectamine2000 according to manufacturer’s instructions. After 24 hours, the media was replaced with fresh media. The next day, the cells were re-seeded at 50% confluency and selected with 10ug/mL Blasticidin and 600ug/mL G418 for 7 days with media change every 2 days. The remaining cells were expanded and tested for reporter construct expression. Translation suppression assay Exemplary TREMs were synthesized and characterized as described herein, then transfected into cells. Forty-eight hours after TREM delivery into cells, conditioned media was collected, fresh media was added to the cells and allowed to equilibrate to room temperature. To measure the efficacy of arginine TREMs in PTC readthrough, full-length HiBiT-tagged disease reporter protein was assayed in both cells, and 48-hour conditioned media. Briefly, reconstituted Nano-Glo® HiBiT Lytic Reagent was added to both cells containing fresh media and 48-hour conditioned media at a 1:1 v/v ratio, mixed on an orbital shaker at 500rpm for 10 minutes, and incubated at room temperature for 10 minutes. The HiBiT-tagged disease reporter activity is measured by reading the luminescence in a plate reader. The data is binned and quantified according to the scheme provided in Example 1.

Claims

Attorney Docket No.: F2099-7037WO What is claimed is: 1. A tRNA effector molecule (TREM) comprising a sequence of Formula (A): [L1]x-[ASt Domain1]-[L2]x-[DH Domain]-[L3]x-[ACH Domain] -[VL Domain]-[TH Domain]-[L4]x-[ASt Domain2]-[L5]x (A), wherein: independently, [L1], [L5], and [VL Domain], are optional; x is 0 or 1; a nucleotide within the TREM sequence comprises a locked nucleic acid (LNA) moiety or 2’,5’-linked nucleotide. 2. The TREM of claim 1, wherein the LNA moiety comprises a heterocyclyl ring or cycloalkyl ring between the 2’-position and the 4’-position of the nucleotide sugar within the nucleotide. 3. The TREM of any one of the preceding claims, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II):
Figure imgf000383_0001
tautomer, or hydrate thereof, wherein: B is a nucleobase; Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of L and X is independently C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each Attorney Docket No.: F2099-7037WO alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of M and Z is absent, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, S(RD)x, -Se(RE)y, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM; provided that no more than two of L, M, Z, and Z may be -O-, -N(RB)-, S(RD)x, -Se(RE)y, and when one of L, M, Z, and Z is O-, -N(RB)-, S(RD)x, -Se(RE)y, the direct covalent binding partner of each selected from L, M, Z, and X is also not O-, -N(RB)-, S(RD)x, -Se(RE)y. 4. The TREM of claim 3, wherein B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). Attorney Docket No.: F2099-7037WO 5. The TREM of any one of the preceding claims, wherein the TREM further comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). 6. The TREM of claim 5, wherein the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. 7. The TREM of any one of claims 5-6, wherein the chemical modification comprises a 2’OMe modification, 2’halo (e.g., 2’F or 2’Cl), modification, 2’MOE modification, 2’-deoxy modification, or a phosphorothioate modification. 8. The TREM of any one of claims 5-7, wherein the TREM does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). 9. The TREM of any one of the preceding claims, wherein TREM comprises a single LNA moiety. 10. The TREM of any one of the preceding claims, wherein the TREM comprises a plurality of LNA moieties. 11. The TREM of any one of the preceding claims, wherein the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 LNA moieties. 12. The TREM of any one of the preceding claims, wherein the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. 13. The TREM of any one of the preceding claims, wherein the LNA moiety is present in the [L1]-[ASt Domain1]-[L2]. Attorney Docket No.: F2099-7037WO 14. The TREM of any one of the preceding claims, wherein the LNA moiety is present in the [L2]-[DH Domain]-[L3]. 15. The TREM of any one of the preceding claims, wherein the LNA moiety is present in the [L3]-[ACH Domain]. 16. The TREM of any one of the preceding claims, wherein the LNA moiety is present in the [VL Domain]. 17. The TREM of any one of the preceding claims, wherein the LNA moiety is present in the [TH Domain]. 18. The TREM of any one of the preceding claims, wherein the LNA moiety is present in the [L4]-[ASt Domain2]-[L5]. 19. The TREM of any one of the preceding claims, wherein the TREM has a sequence selected from a sequence provided in FIG.1. 20. The TREM of any one of the preceding claims, wherein the TREM comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in FIG.1. 21. The TREM of any one of the preceding claims, wherein the TREM comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from a TREM provided in FIG.1. 22. The TREM of any one of the preceding claims, wherein the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’-MOE, 2’-deoxy, LNA, or phosphorothiorate modifications). Attorney Docket No.: F2099-7037WO 23. The TREM of any one of the preceding claims, wherein the TREM comprises the nucleotide sequence of any one of SEQ ID NOs: 625-1416. 24. The TREM of any one of the preceding claims, wherein the nucleotide within the TREM sequence comprising the LNA moiety has a structure of Formula (II-e): or hydrate thereof, wherein:
Figure imgf000387_0001
B is a nucleobase; Y is C(R’), O, or N(RB); each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; X is C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C1-C6 heteroalkylene C1-C6 haloalkylene -O-, -N(RB)-, -S(RD)x-, -Se(RE)y-, wherein each alkylene, alkenylene, alkynylene, heteroalkylene, and haloalkylene is optionally substituted with one or more R11; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or Attorney Docket No.: F2099-7037WO RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; n is 1, 2, 3, 4, 5, or 6; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 25. The TREM of any one of the preceding claims, wherein the nucleotide within the TREM sequencing comprising the LNA moiety has a structure of Formula (II-m): ,
Figure imgf000388_0001
or hydrate thereof, wherein: B is a nucleobase; each of R1, R2, and R3 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R4a and R4b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; Attorney Docket No.: F2099-7037WO each of R6a and R6b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; each RD and RE is independently C1-C6 alkyl or oxo; each of R10 and R11 is independently C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 26. The TREM of any one of the preceding claims, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III):
Figure imgf000389_0001
tautomer, or hydrate thereof, wherein: B is a nucleobase; Y is C(R’), O, or N(RB); each of R1, R2, and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; Attorney Docket No.: F2099-7037WO each of R3a and R3b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 27. The TREM of claim 26, wherein B is a naturally occurring nucleobase (e.g., adenine, cytosine, guanine, uracil). 28. The TREM of any one of claims 26-27, wherein the TREM further comprises a chemical modification (e.g., a naturally occurring chemical modification or a non-naturally occurring chemical modification). 29. The TREM of any one of claims 26-28, wherein the chemical modification is a nucleotide sugar modification, a nucleobase modification, or an internucleotide linkage modification. Attorney Docket No.: F2099-7037WO 30. The TREM of any one of claims 28-29, wherein the chemical modification comprises a 2’OMe modification, 2’halo (e.g., 2’F or 2’Cl) modification, 2’MOE modification, 2’-deoxy modification, or phosphorothioate modification. 31. The TREM of any one of claims 26-30, wherein the compound of Formula (III) does not comprise a chemical modification (e.g., a naturally occurring chemical modification or a non- naturally occurring chemical modification). 32. The TREM of any one of claims 26-31, wherein TREM comprises a single 2’,5’-linked nucleotide moiety. 33. The TREM of any one of claims 26-32, wherein the TREM comprises a plurality of 2’,5’-linked nucleotide moieties. 34. The TREM of any one of claims 26-33, wherein the TREM comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 252’,5’-linked nucleotides. 35. The TREM of any one of claims 26-34, wherein the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]-[L2]. 36. The TREM of any one of claims 26-35, wherein the 2’,5’-linked nucleotide is present in the [L1]-[ASt Domain1]-[L2]. 37. The TREM of any one of claims 26-36, wherein the 2’,5’-linked nucleotide is present in the [L2]-[DH Domain]-[L3]. 38. The TREM of any one of claims 26-37, wherein the 2’,5’-linked nucleotide is present in the [L3]-[ACH Domain]. 39. The TREM of any one of claims 26-38, wherein the 2’,5’-linked nucleotide is present in the [VL Domain]. Attorney Docket No.: F2099-7037WO 40. The TREM of any one of claims 26-39, wherein the 2’,5’-linked nucleotide is present in the [TH Domain]. 41. The TREM of any one of claims 26-40, wherein the 2’,5’-linked nucleotide is present in the [L4]-[ASt Domain2]-[L5]. 42. The TREM of any one of claims 26-41, wherein the TREM has a sequence selected from a sequence provided in FIG.1. 43. The TREM of any one of claims 26-42, wherein the TREM comprises a TREM having at least 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with a TREM provided in FIG. 1. 44. The TREM of any one of claims 26-43, wherein the TREM comprises a sequence that differs by no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides from a TREM provided in FIG. 1. 45. The TREM of any one of claims 26-44, wherein the TREM comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional non-naturally occurring modifications compared with a TREM provided in FIG.1 (e.g., 2’-ribose modifications or an internucleotide modification, e.g., 2’OMe, 2’-halo, 2’- MOE, 2’-deoxy, or phosphorothiorate modifications). 46. The TREM of any one of claims 26-46, wherein the nucleotide within the TREM sequence comprising the 2’,5’-linked nucleotide has a structure of Formula (III-a):
Figure imgf000392_0001
or hydrate thereof, wherein: Attorney Docket No.: F2099-7037WO B is a nucleobase; Y is C(R’), O, or N(RB); each of R1, R2, and R4 is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R3a is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R3c is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, or C1-C6 haloalkyl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; each of R5a and R5b is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, -ORA, or -N(RB)(RC), wherein each alkyl, alkenyl, alkynyl, heteroalkyl, and haloalkyl is optionally substituted with one or more R10; R’ is hydrogen or C1-C6 alkyl; each RA is independently hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl; each of RB and RC is independently hydrogen, C1-C6 alkyl, C1-C6-heteroalkyl, cycloalkyl, or heterocyclyl; or RB and RC together with the atom to which they are attached form a 3-7-membered heterocyclyl ring optionally substituted with one or more R11; R10 is C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 haloalkyl, halo, cyano, or -ORA1; RA1 is hydrogen or C1-C6 alkyl; each of x and y is independently 0, 1, or 2; and “ ” refers to an attachment point to the rest of the TREM sequence, another atom, or the terminus of the TREM. 47. The TREM of any one of the preceding claims, wherein the TREM comprises the nucleotide sequence of any one of SEQ ID NOs: 625-793. Attorney Docket No.: F2099-7037WO 48. The TREM of any one of the preceding claims, wherein the TREM comprises the nucleotide sequence of any one of SEQ ID NOs: 793-1416. 49. The TREM of any one of claims 26-45, wherein the TREM comprises a nucleotide sequence selected from any one of SEQ ID NOs: 625-1416. 50. A pharmaceutical composition comprising a TREM of any one of the preceding claims. 51. The pharmaceutical composition of claim 50, further comprising a pharmaceutically acceptable component, e.g., an excipient. 52. A lipid nanoparticle formulation comprising a TREM of any one of claims 1-49. 53. A lipid nanoparticle formulation comprising a pharmaceutical composition of claims 50 or 51. 54. A composition for use in treating a subject having a disease or disorder associated with a PTC comprising administering to the subject a TREM described herein (e.g., a TREM of any one of claims 1-49). 55. The composition for use of claim 54, wherein the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease. 56. A method of treating a subject having a disease or disorder associated with a PTC comprising administering to the subject a TREM described herein (e.g., a TREM of any one of claims 1-49), thereby treating the subject having the disease or disorder. 57. The method of claim 56, wherein the disease or disorder associated with a PTC comprises Hemophilia B, Fabry disease, Usher syndrome, or CLN2 disease.
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