CA3024184A1 - Variants of a dna polymerase of the polx family - Google Patents
Variants of a dna polymerase of the polx family Download PDFInfo
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- CA3024184A1 CA3024184A1 CA3024184A CA3024184A CA3024184A1 CA 3024184 A1 CA3024184 A1 CA 3024184A1 CA 3024184 A CA3024184 A CA 3024184A CA 3024184 A CA3024184 A CA 3024184A CA 3024184 A1 CA3024184 A1 CA 3024184A1
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- variant
- residue
- dna polymerase
- seq
- nucleic acid
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Classifications
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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
- C12N15/63—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
-
- 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)
- C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
-
- 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
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR1655475A FR3052462A1 (fr) | 2016-06-14 | 2016-06-14 | Variants d'une adn polymerase de la famille polx |
| FR1655475 | 2016-06-14 | ||
| PCT/FR2017/051519 WO2017216472A2 (fr) | 2016-06-14 | 2017-06-13 | Variants d'une adn polymérase de la famille polx |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA3024184A1 true CA3024184A1 (en) | 2017-12-21 |
Family
ID=57348782
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA3024184A Pending CA3024184A1 (en) | 2016-06-14 | 2017-06-13 | Variants of a dna polymerase of the polx family |
Country Status (11)
| Country | Link |
|---|---|
| US (3) | US20200002690A1 (OSRAM) |
| EP (1) | EP3469075A2 (OSRAM) |
| JP (2) | JP7118510B2 (OSRAM) |
| KR (1) | KR102522263B1 (OSRAM) |
| CN (1) | CN109477080A (OSRAM) |
| AU (2) | AU2017286477C1 (OSRAM) |
| CA (1) | CA3024184A1 (OSRAM) |
| FR (2) | FR3052462A1 (OSRAM) |
| IL (1) | IL263503B2 (OSRAM) |
| SG (2) | SG11201809961TA (OSRAM) |
| WO (1) | WO2017216472A2 (OSRAM) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020099451A1 (en) * | 2018-11-14 | 2020-05-22 | Dna Script | Terminal deoxynucleotidyl transferase variants and uses thereof |
| WO2021116270A1 (en) * | 2019-12-12 | 2021-06-17 | Dna Script | Chimeric terminal deoxynucleotidyl transferases for template-free enzymatic synthesis of polynucleotides |
| JP2022513371A (ja) * | 2018-10-19 | 2022-02-07 | モレキュラー アッセンブリーズ， インコーポレイテッド | ポリデオキシヌクレオチド合成のための改変テンプレート非依存性酵素 |
| JP2023528477A (ja) * | 2020-06-03 | 2023-07-04 | モレキュラー アッセンブリーズ， インコーポレイテッド | 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 |
| EP4653525A1 (en) * | 2024-05-21 | 2025-11-26 | Uab "Biomatter Designs" | Engineered terminal deoxynucleotidyl transferase polymerases |
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|---|---|---|---|---|
| GB2577020B (en) | 2017-05-26 | 2020-09-16 | Nuclera Nucleics Ltd | Use of terminal transferase enzyme in nucleic acid synthesis |
| EP3665274B1 (en) | 2017-08-07 | 2025-10-15 | DNA Script | Variants of family a dna polymerase and uses thereof |
| CA3087714A1 (en) * | 2018-01-08 | 2019-07-11 | Dna Script | Variants of terminal deoxynucleotidyl transferase and uses thereof |
| US10752887B2 (en) | 2018-01-08 | 2020-08-25 | Dna Script | Variants of terminal deoxynucleotidyl transferase and uses thereof |
| EP3826762A1 (en) | 2018-07-23 | 2021-06-02 | DNA Script | Massively parallel enzymatic synthesis of nucleic acid strands |
| AU2019398307A1 (en) | 2018-12-13 | 2021-06-10 | Dna Script | Direct oligonucleotide synthesis on cells and biomolecules |
| JP7491934B2 (ja) | 2019-01-03 | 2024-05-28 | ディーエヌエー スクリプト | オリゴヌクレオチドセットのワンポット合成 |
| GB201901501D0 (en) | 2019-02-04 | 2019-03-27 | Nuclera Nucleics Ltd | Modified terminal deoxynucleotidyl transferase (TdT) enzymes |
| US11359221B2 (en) | 2019-02-12 | 2022-06-14 | Dna Script Sas | Efficient product cleavage in template-free enzymatic synthesis of polynucleotides |
| EP3744854A1 (en) * | 2019-05-28 | 2020-12-02 | DNA Script | Variants of terminal deoxynucleotidyl transferase and uses thereof |
| JP2022534004A (ja) * | 2019-05-28 | 2022-07-27 | ディーエヌエー スクリプト | 末端デオキシヌクレオチジルトランスフェラーゼの変異体及びその使用 |
| WO2021018919A1 (en) | 2019-07-30 | 2021-02-04 | Dna Script | Template-free enzymatic synthesis of polynucleotides using poly(a) and poly(u) polymerases |
| US20220403435A1 (en) | 2019-08-01 | 2022-12-22 | Dna Script Sas | Increasing Long-Sequence Yields In Template-Free Enzymatic Synthesis of Polynucleotides |
| CN110331136B (zh) * | 2019-09-05 | 2019-12-24 | 中国科学院天津工业生物技术研究所 | 一种末端脱氧核糖核苷转移酶变体及其应用 |
| WO2021048142A1 (en) | 2019-09-09 | 2021-03-18 | Dna Script | Template-free enzymatic polynucleotide synthesis using photocleavable linkages |
| EP4034669A1 (en) | 2019-09-23 | 2022-08-03 | DNA Script | Increasing long-sequence yields in template-free enzymatic synthesis of polynucleotides |
| KR20220097976A (ko) | 2019-11-13 | 2022-07-08 | 디엔에이 스크립트 | 폴리뉴클레오타이드의 무-주형 고 효율 효소 합성 |
| US20230203553A1 (en) | 2019-12-16 | 2023-06-29 | Dna Script | Template-Free Enzymatic Polynucleotide Synthesis Using Dismutationless Terminal Deoxynucleotidyl Transferase Variants |
| CN115151652B (zh) | 2020-02-25 | 2025-02-07 | Dna斯克瑞普特公司 | 用于酶促合成多核苷酸的方法和设备 |
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| AU2021271831A1 (en) * | 2020-05-12 | 2022-12-08 | National University Of Singapore | Thermostable terminal deoxynucleotidyl transferase |
| US12209266B2 (en) | 2020-05-12 | 2025-01-28 | Illumina, Inc. | Generating nucleic acids with modified bases using recombinant terminal deoxynucleotidyl transferase |
| WO2021250265A2 (en) | 2020-06-12 | 2021-12-16 | Synhelix | Ab-initio, template-independent synthesis of nucleic acids using thermostable enzymes |
| CN115803108A (zh) | 2020-06-16 | 2023-03-14 | Dna斯克瑞普特公司 | 用于酶促多核苷酸合成的系统、设备和套件 |
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| KR20230041975A (ko) | 2020-07-21 | 2023-03-27 | 일루미나, 싱가포르 피티이 엘티디 | Tdt-기반 효소적 핵산을 위한 기질로서의 염기-변형된 뉴클레오티드 |
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| CN120530202A (zh) | 2023-01-26 | 2025-08-22 | Dna斯克瑞普特公司 | 多核苷酸探针的酶促合成 |
| CN121039286A (zh) | 2023-05-03 | 2025-11-28 | Dna斯克瑞普特公司 | 用于3'-o-氨基多核苷酸的去保护的试剂 |
| WO2024241324A1 (en) * | 2023-05-24 | 2024-11-28 | Scintium Ltd | Method for training an artificial intelligence (ai) model and generating a 3d computer-aided design (cad) and system therefor |
| WO2024256604A1 (en) | 2023-06-15 | 2024-12-19 | Dna Script | Salt addition during enzymatic polynucleotide synthesis |
| WO2025003434A1 (en) | 2023-06-30 | 2025-01-02 | Dna Script | Nucleic acid synthesis on reusable support |
| WO2025021984A1 (en) | 2023-07-27 | 2025-01-30 | Dna Script | Method for generating a plurality of polynucleotides |
| WO2025088024A1 (en) | 2023-10-24 | 2025-05-01 | Dna Script | Novel terminal deoxynucleotidyl transferase (tdt) variants and uses thereof |
| WO2025093659A1 (en) | 2023-11-04 | 2025-05-08 | Dna Script | 3'-o-aminooxymethyl blocked nucleotides and use thereof in enzymatic synthesis of polynucleotides |
| WO2025137872A1 (zh) * | 2023-12-26 | 2025-07-03 | 华大工程生物学长荡湖研究所 | 一种末端脱氧核糖核苷转移酶突变体及其制备方法和应用 |
| WO2025155239A1 (en) * | 2024-01-17 | 2025-07-24 | Agency For Science, Technology And Research | X family dna polymerases and uses thereof |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5004689A (en) | 1982-02-22 | 1991-04-02 | Biogen, Massachusetts | DNA sequences, recombinant DNA molecules and processes for producing human gamma interferon-like polypeptides in high yields |
| IL71691A (en) | 1984-04-27 | 1991-04-15 | Yeda Res & Dev | Production of interferon-ypsilon |
| US5047335A (en) | 1988-12-21 | 1991-09-10 | The Regents Of The University Of Calif. | Process for controlling intracellular glycosylation of proteins |
| BG52073B2 (en) | 1990-01-24 | 1996-04-30 | Inst Molekuljarna Biolog | Method for the preparation of recombinant human noncystein -interferon, free of n-end methionine |
| US6812339B1 (en) * | 2000-09-08 | 2004-11-02 | Applera Corporation | Polymorphisms in known genes associated with human disease, methods of detection and uses thereof |
| EP2352818B1 (en) * | 2008-11-03 | 2016-02-10 | Kapa Biosystems, Inc. | Chimeric dna polymerases |
| WO2012119095A1 (en) * | 2011-03-02 | 2012-09-07 | Board Of Regents, The University Of Texas System | Fus1/tusc2 therapies |
| EP3080158A4 (en) * | 2013-12-09 | 2018-03-21 | Adimab, LLC | Polyclonal mixtures of antibodies, and methods of making and using them |
| FR3020071B1 (fr) | 2014-04-17 | 2017-12-22 | Dna Script | Procede de synthese d'acides nucleiques, notamment d'acides nucleiques de grande longueur, utilisation du procede et kit pour la mise en œuvre du procede |
| FR3025201B1 (fr) | 2014-09-02 | 2018-10-12 | Dna Script | Nucleotides modifies pour la synthese d'acides nucleiques, un kit renfermant de tels nucleotides et leur utilisation pour la production de genes ou sequences d'acides nucleiques synthetiques |
| US10059929B2 (en) * | 2014-10-20 | 2018-08-28 | Molecular Assemblies, Inc. | Modified template-independent enzymes for polydeoxynucleotide synthesis |
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2023
- 2023-11-21 US US18/516,616 patent/US20240124855A1/en active Pending
- 2023-12-13 AU AU2023282219A patent/AU2023282219A1/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2022513371A (ja) * | 2018-10-19 | 2022-02-07 | モレキュラー アッセンブリーズ， インコーポレイテッド | ポリデオキシヌクレオチド合成のための改変テンプレート非依存性酵素 |
| EP3867361A4 (en) * | 2018-10-19 | 2022-07-27 | Molecular Assemblies, Inc. | MODIFIED TEMPLATE-INDEPENDENT ENZYMES FOR POLYDEOXYNUCLEOTIDE SYNTHESIS |
| WO2020099451A1 (en) * | 2018-11-14 | 2020-05-22 | Dna Script | Terminal deoxynucleotidyl transferase variants and uses thereof |
| CN113302289A (zh) * | 2018-11-14 | 2021-08-24 | Dna斯克瑞普特公司 | 末端脱氧核苷酸转移酶变体及其用途 |
| US11859217B2 (en) | 2018-11-14 | 2024-01-02 | Dna Script | Terminal deoxynucleotidyl transferase variants and uses thereof |
| US12371680B2 (en) | 2018-11-14 | 2025-07-29 | Dna Script Sas | Terminal deoxynucleotidyl transferase variants and uses thereof |
| WO2021116270A1 (en) * | 2019-12-12 | 2021-06-17 | Dna Script | Chimeric terminal deoxynucleotidyl transferases for template-free enzymatic synthesis of polynucleotides |
| JP2023528477A (ja) * | 2020-06-03 | 2023-07-04 | モレキュラー アッセンブリーズ， インコーポレイテッド | 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 |
| EP4653525A1 (en) * | 2024-05-21 | 2025-11-26 | Uab "Biomatter Designs" | Engineered terminal deoxynucleotidyl transferase polymerases |
| WO2025243149A1 (en) * | 2024-05-21 | 2025-11-27 | Uab Biomatter Designs | Engineered terminal deoxynucleotidyl transferase polymerases |
Also Published As
| Publication number | Publication date |
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| JP2019517799A (ja) | 2019-06-27 |
| SG11201809961TA (en) | 2018-12-28 |
| CN109477080A (zh) | 2019-03-15 |
| US20240124855A1 (en) | 2024-04-18 |
| IL263503B2 (en) | 2025-04-01 |
| AU2017286477A1 (en) | 2018-12-06 |
| WO2017216472A2 (fr) | 2017-12-21 |
| JP2022137127A (ja) | 2022-09-21 |
| KR102522263B1 (ko) | 2023-04-18 |
| WO2017216472A3 (fr) | 2018-03-15 |
| US20230167421A1 (en) | 2023-06-01 |
| AU2017286477C1 (en) | 2023-12-21 |
| FR3053699B1 (fr) | 2021-04-02 |
| FR3053699A1 (fr) | 2018-01-12 |
| AU2017286477B2 (en) | 2023-09-14 |
| FR3052462A1 (fr) | 2017-12-15 |
| IL263503B1 (en) | 2024-12-01 |
| AU2023282219A1 (en) | 2024-01-18 |
| IL263503A (en) | 2019-02-03 |
| US20200002690A1 (en) | 2020-01-02 |
| JP7118510B2 (ja) | 2022-08-16 |
| KR20190024883A (ko) | 2019-03-08 |
| EP3469075A2 (fr) | 2019-04-17 |
| SG10202009688QA (en) | 2020-11-27 |
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