WO2021247851A2 - Enzymes indépendantes d'une matrice modifiée pour la synthèse de polydésoxynucléotides - Google Patents

Enzymes indépendantes d'une matrice modifiée pour la synthèse de polydésoxynucléotides Download PDF

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Publication number
WO2021247851A2
WO2021247851A2 PCT/US2021/035685 US2021035685W WO2021247851A2 WO 2021247851 A2 WO2021247851 A2 WO 2021247851A2 US 2021035685 W US2021035685 W US 2021035685W WO 2021247851 A2 WO2021247851 A2 WO 2021247851A2
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WIPO (PCT)
Prior art keywords
tdt
modified
modified tdt
mutations
lys
Prior art date
Application number
PCT/US2021/035685
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English (en)
Other versions
WO2021247851A3 (fr
Inventor
Julie L. TUBBS
Prem SINHA
Boguslaw STEC
Matthew T. HOLDEN
Christopher Wilson
J. William Efcavitch
Deanne W. SAMMOND
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 CN202180057936.6A priority Critical patent/CN116075593A/zh
Priority to CA3185851A priority patent/CA3185851A1/fr
Priority to JP2022574577A priority patent/JP2023528477A/ja
Priority to KR1020237000176A priority patent/KR20240016237A/ko
Priority to EP21818476.0A priority patent/EP4162035A2/fr
Priority to AU2021285906A priority patent/AU2021285906A1/en
Publication of WO2021247851A2 publication Critical patent/WO2021247851A2/fr
Publication of WO2021247851A3 publication Critical patent/WO2021247851A3/fr

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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)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Biophysics (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

La présente invention concerne des procédés permettant d'identifier des polymérases, telles que des nucléotidyl-transférases terminales (TdT) modifiées, qui sont capables de lier des nucléotides comprenant des fragments de blocage 3'-O- séparables à un initiateur d'acide nucléique, sans utiliser de matrice. L'invention comprend en outre les polymérases identifiées, et des procédés d'utilisation desdites polymérases pour la synthèse de novo de séquences d'oligonucléotides prédéfinies.
PCT/US2021/035685 2020-06-03 2021-06-03 Enzymes indépendantes d'une matrice modifiée pour la synthèse de polydésoxynucléotides WO2021247851A2 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
CN202180057936.6A CN116075593A (zh) 2020-06-03 2021-06-03 用于聚脱氧核苷酸合成的经修饰的非模板依赖性酶
CA3185851A CA3185851A1 (fr) 2020-06-03 2021-06-03 Enzymes independantes d'une matrice modifiee pour la synthese de polydesoxynucleotides
JP2022574577A JP2023528477A (ja) 2020-06-03 2021-06-03 ポリデオキシヌクレオチド合成のための改変された鋳型非依存的酵素
KR1020237000176A KR20240016237A (ko) 2020-06-03 2021-06-03 폴리데옥시뉴클레오티드 합성을 위한 변형된 템플레이트-비의존성 효소
EP21818476.0A EP4162035A2 (fr) 2020-06-03 2021-06-03 Enzymes indépendantes d'une matrice modifiée pour la synthèse de polydésoxynucléotides
AU2021285906A AU2021285906A1 (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 true WO2021247851A2 (fr) 2021-12-09
WO2021247851A3 WO2021247851A3 (fr) 2022-03-17

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2021/035685 WO2021247851A2 (fr) 2020-06-03 2021-06-03 Enzymes indépendantes d'une matrice modifiée pour la synthèse de polydésoxynucléotides

Country Status (7)

Country Link
EP (1) EP4162035A2 (fr)
JP (1) JP2023528477A (fr)
KR (1) KR20240016237A (fr)
CN (1) CN116075593A (fr)
AU (1) AU2021285906A1 (fr)
CA (1) CA3185851A1 (fr)
WO (1) WO2021247851A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114410602A (zh) * 2022-01-28 2022-04-29 赛纳生物科技(北京)有限公司 一种末端脱氧核苷酸转移酶的突变体及其应用

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2496875C2 (ru) * 2007-03-12 2013-10-27 ДАНИСКО ЮЭс ИНК. Мoдифицированные протеазы
EP3699283A1 (fr) * 2014-10-20 2020-08-26 Molecular Assemblies Inc. Enzymes indépendantes de la matrice modifiées pour la synthèse de polydésoxynucléotides
GB201502152D0 (en) * 2015-02-10 2015-03-25 Nuclera Nucleics Ltd Novel use
US10435676B2 (en) * 2018-01-08 2019-10-08 Dna Script Variants of terminal deoxynucleotidyl transferase and uses thereof

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114410602A (zh) * 2022-01-28 2022-04-29 赛纳生物科技(北京)有限公司 一种末端脱氧核苷酸转移酶的突变体及其应用
CN114410602B (zh) * 2022-01-28 2024-01-19 赛纳生物科技(北京)有限公司 一种末端脱氧核苷酸转移酶的突变体及其应用

Also Published As

Publication number Publication date
JP2023528477A (ja) 2023-07-04
CA3185851A1 (fr) 2021-12-09
WO2021247851A3 (fr) 2022-03-17
KR20240016237A (ko) 2024-02-06
EP4162035A2 (fr) 2023-04-12
AU2021285906A1 (en) 2023-02-02
CN116075593A (zh) 2023-05-05

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