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WO2021247851A3 - Modified template-independent enzymes for polydeoxynucleotide synthesis - Google Patents

Modified template-independent enzymes for polydeoxynucleotide synthesis Download PDF

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Publication number
WO2021247851A3
WO2021247851A3 PCT/US2021/035685 US2021035685W WO2021247851A3 WO 2021247851 A3 WO2021247851 A3 WO 2021247851A3 US 2021035685 W US2021035685 W US 2021035685W WO 2021247851 A3 WO2021247851 A3 WO 2021247851A3
Authority
WO
WIPO (PCT)
Prior art keywords
polymerases
modified template
independent enzymes
polydeoxynucleotide synthesis
synthesis
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2021/035685
Other languages
French (fr)
Other versions
WO2021247851A2 (en
Inventor
Julie L. TUBBS
Prem SINHA
Boguslaw STEC
Matthew T. HOLDEN
Christopher Wilson
J. William Efcavitch
Deanne W. SAMMOND
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Molecular Assemblies Inc
Original Assignee
Molecular Assemblies Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US16/891,449 external-priority patent/US11390858B2/en
Application filed by Molecular Assemblies Inc filed Critical Molecular Assemblies Inc
Priority to EP21818476.0A priority Critical patent/EP4162035A4/en
Priority to JP2022574577A priority patent/JP2023528477A/en
Priority to CN202180057936.6A priority patent/CN116075593A/en
Priority to AU2021285906A priority patent/AU2021285906A1/en
Priority to KR1020237000176A priority patent/KR20240016237A/en
Priority to CA3185851A priority patent/CA3185851A1/en
Publication of WO2021247851A2 publication Critical patent/WO2021247851A2/en
Publication of WO2021247851A3 publication Critical patent/WO2021247851A3/en
Anticipated expiration legal-status Critical
Ceased 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • C12N9/1264—DNA nucleotidylexotransferase (2.7.7.31), i.e. terminal nucleotidyl transferase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/07—Nucleotidyltransferases (2.7.7)
    • C12Y207/07031—DNA nucleotidylexotransferase (2.7.7.31), i.e. terminal deoxynucleotidyl transferase
    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Saccharide Compounds (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention includes methods for identifying polymerases, such as modified terminal nucleotidyl transferases (TdT), that are capable of binding nucleotides comprising removable 3'- O-blocking moieties to a nucleic acid initiator, without the use of a template. The invention further includes the identified polymerases, and methods of using the polymerases for de novo synthesis of predetermined oligonucleotide sequences.
PCT/US2021/035685 2020-06-03 2021-06-03 Modified template-independent enzymes for polydeoxynucleotide synthesis Ceased WO2021247851A2 (en)

Priority Applications (6)

Application Number Priority Date Filing Date Title
EP21818476.0A EP4162035A4 (en) 2020-06-03 2021-06-03 MODIFIED, MATRIX-INDEPENDENT ENZYMES FOR THE SYNTHESIS OF POLYDEOXYNUCLEOTIDES
JP2022574577A JP2023528477A (en) 2020-06-03 2021-06-03 A modified template-independent enzyme for polydeoxynucleotide synthesis
CN202180057936.6A CN116075593A (en) 2020-06-03 2021-06-03 Modified template-independent enzymes for polydeoxynucleotide synthesis
AU2021285906A AU2021285906A1 (en) 2020-06-03 2021-06-03 Modified template-independent enzymes for polydeoxynucleotide synthesis
KR1020237000176A KR20240016237A (en) 2020-06-03 2021-06-03 Modified template-independent enzymes for polydeoxynucleotide synthesis
CA3185851A CA3185851A1 (en) 2020-06-03 2021-06-03 Modified template-independent enzymes for polydeoxynucleotide synthesis

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US16/891,449 US11390858B2 (en) 2014-10-20 2020-06-03 Modified template-independent enzymes for polydeoxynucleotide synthesis
US16/891,449 2020-06-03

Publications (2)

Publication Number Publication Date
WO2021247851A2 WO2021247851A2 (en) 2021-12-09
WO2021247851A3 true WO2021247851A3 (en) 2022-03-17

Family

ID=78831733

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2021/035685 Ceased WO2021247851A2 (en) 2020-06-03 2021-06-03 Modified template-independent enzymes for polydeoxynucleotide synthesis

Country Status (7)

Country Link
EP (1) EP4162035A4 (en)
JP (1) JP2023528477A (en)
KR (1) KR20240016237A (en)
CN (1) CN116075593A (en)
AU (1) AU2021285906A1 (en)
CA (1) CA3185851A1 (en)
WO (1) WO2021247851A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114410602B (en) * 2022-01-28 2024-01-19 赛纳生物科技(北京)有限公司 Mutant of terminal deoxynucleotidyl transferase and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140295526A1 (en) * 2007-03-12 2014-10-02 Danisco Us Inc. Modified proteases
US20160108382A1 (en) * 2014-10-20 2016-04-21 Molecular Assemblies, Inc. Modified template-independent enzymes for polydeoxynucleotide synthesis
US20180023108A1 (en) * 2015-02-10 2018-01-25 Nuclera Nucleics Ltd. Novel use
US20190211315A1 (en) * 2018-01-08 2019-07-11 Dna Script Variants of Terminal Deoxynucleotidyl Transferase and Uses Thereof

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10215035A1 (en) * 2002-04-05 2003-10-23 Roche Diagnostics Gmbh Recombinant terminal deoxynucleotide transferase with improved functionality
US10774316B2 (en) * 2014-10-20 2020-09-15 Molecular Assemblies, Inc. Modified template-independent enzymes for polydeoxynucleotide synthesis
FR3052462A1 (en) * 2016-06-14 2017-12-15 Dna Script POLYMERASE DNA VARIANTS OF THE POLX FAMILY

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140295526A1 (en) * 2007-03-12 2014-10-02 Danisco Us Inc. Modified proteases
US20160108382A1 (en) * 2014-10-20 2016-04-21 Molecular Assemblies, Inc. Modified template-independent enzymes for polydeoxynucleotide synthesis
US20180023108A1 (en) * 2015-02-10 2018-01-25 Nuclera Nucleics Ltd. Novel use
US20190211315A1 (en) * 2018-01-08 2019-07-11 Dna Script Variants of Terminal Deoxynucleotidyl Transferase and Uses Thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
TROSHCHYNSKY, A ET AL.: "Functional analyses of polymorphic variants of human terminal deoxynucleotidyl transferase", GENES AND IMMUNIT Y, vol. 16, no. 6, 4 June 2015 (2015-06-04), pages 388 - 398, XP037161848, [retrieved on 20150900], DOI: 10.1038/ gene .2015.19 *

Also Published As

Publication number Publication date
AU2021285906A1 (en) 2023-02-02
WO2021247851A2 (en) 2021-12-09
EP4162035A2 (en) 2023-04-12
CA3185851A1 (en) 2021-12-09
EP4162035A4 (en) 2025-03-12
JP2023528477A (en) 2023-07-04
CN116075593A (en) 2023-05-05
KR20240016237A (en) 2024-02-06

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