CA2978194C - Polymerase variants and uses thereof - Google Patents

Polymerase variants and uses thereof Download PDF

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Publication number
CA2978194C
CA2978194C CA2978194A CA2978194A CA2978194C CA 2978194 C CA2978194 C CA 2978194C CA 2978194 A CA2978194 A CA 2978194A CA 2978194 A CA2978194 A CA 2978194A CA 2978194 C CA2978194 C CA 2978194C
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CA
Canada
Prior art keywords
polymerase
dna
modified
dna polymerase
nucleotide
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.)
Active
Application number
CA2978194A
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English (en)
French (fr)
Other versions
CA2978194A1 (en
Inventor
Aruna Ayer
Cleoma Arnold
Charles Schwab
Eileen Thai
Ilya Lederman
Colin Mcgaw
Tyler Shultz
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.)
F Hoffmann La Roche AG
Original Assignee
F Hoffmann La Roche AG
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 US15/012,317 external-priority patent/US10308918B2/en
Application filed by F Hoffmann La Roche AG filed Critical F Hoffmann La Roche AG
Priority to CA3028797A priority Critical patent/CA3028797C/en
Publication of CA2978194A1 publication Critical patent/CA2978194A1/en
Application granted granted Critical
Publication of CA2978194C publication Critical patent/CA2978194C/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • 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)
    • 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
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/07—Nucleotidyltransferases (2.7.7)
    • 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)
    • C12Y207/07007—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
CA2978194A 2015-05-14 2016-05-13 Polymerase variants and uses thereof Active CA2978194C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CA3028797A CA3028797C (en) 2015-05-14 2016-05-13 Polymerase variants and uses thereof

Applications Claiming Priority (9)

Application Number Priority Date Filing Date Title
US201562161571P 2015-05-14 2015-05-14
US62/161,571 2015-05-14
US201562202895P 2015-08-09 2015-08-09
US62/202,895 2015-08-09
US15/012,317 US10308918B2 (en) 2015-02-02 2016-02-01 Polymerase variants
US15/012,317 2016-02-01
US15/151,364 US10526588B2 (en) 2015-05-14 2016-05-10 Polymerase variants and uses thereof
US15/151,364 2016-05-10
PCT/US2016/032258 WO2016183403A2 (en) 2015-05-14 2016-05-13 Polymerase variants and uses thereof

Related Child Applications (1)

Application Number Title Priority Date Filing Date
CA3028797A Division CA3028797C (en) 2015-05-14 2016-05-13 Polymerase variants and uses thereof

Publications (2)

Publication Number Publication Date
CA2978194A1 CA2978194A1 (en) 2016-11-17
CA2978194C true CA2978194C (en) 2021-11-16

Family

ID=57248698

Family Applications (2)

Application Number Title Priority Date Filing Date
CA2978194A Active CA2978194C (en) 2015-05-14 2016-05-13 Polymerase variants and uses thereof
CA3028797A Active CA3028797C (en) 2015-05-14 2016-05-13 Polymerase variants and uses thereof

Family Applications After (1)

Application Number Title Priority Date Filing Date
CA3028797A Active CA3028797C (en) 2015-05-14 2016-05-13 Polymerase variants and uses thereof

Country Status (6)

Country Link
US (3) US10526588B2 (https=)
EP (1) EP3294749A4 (https=)
JP (3) JP2018515103A (https=)
CN (1) CN107636004A (https=)
CA (2) CA2978194C (https=)
WO (1) WO2016183403A2 (https=)

Families Citing this family (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6736565B2 (ja) 2015-02-02 2020-08-05 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ ポリメラーゼバリアント
US10526588B2 (en) * 2015-05-14 2020-01-07 Roche Sequencing Solutions, Inc. Polymerase variants and uses thereof
US10745685B2 (en) 2015-11-16 2020-08-18 Stratos Genomics, Inc. DP04 polymerase variants
EP3423574B1 (en) * 2016-02-29 2021-03-17 Genia Technologies, Inc. Polymerase-template complexes for nanopore sequencing
CN108699539B (zh) 2016-02-29 2022-11-18 吉尼亚科技公司 外切核酸酶缺陷的聚合酶
US10590480B2 (en) 2016-02-29 2020-03-17 Roche Sequencing Solutions, Inc. Polymerase variants
US10655174B2 (en) 2016-05-27 2020-05-19 Roche Sequencing Solutions, Inc. Tagged multi-nucleotides useful for nucleic acid sequencing
US10669580B2 (en) 2016-08-26 2020-06-02 Roche Sequencing Solutions, Inc. Tagged nucleotides useful for nanopore detection
US10829747B2 (en) 2016-09-22 2020-11-10 Roche Sequencing Solutions, Inc. Polymerase variants
WO2018204707A1 (en) 2017-05-04 2018-11-08 Stratos Genomics Inc. Dp04 polymerase variants
CA3084812A1 (en) 2017-12-11 2019-06-20 Stratos Genomics, Inc. Dpo4 polymerase variants with improved accuracy
WO2019166457A1 (en) 2018-02-28 2019-09-06 F. Hoffmann-La Roche Ag Tagged nucleoside compounds useful for nanopore detection
CN112041331B (zh) 2018-02-28 2024-05-28 豪夫迈·罗氏有限公司 α-溶血素变体及其用途
WO2021156370A1 (en) 2020-02-06 2021-08-12 F. Hoffmann-La Roche Ag Compositions that reduce template threading into a nanopore
EP4196608B1 (en) 2020-08-11 2024-10-23 F. Hoffmann-La Roche AG Nucleoside-5'-oligophosphates tagged with positively-charged polymers, nanopores incorporating negative charges, and methods and systems using the same
EP4355757A1 (en) 2021-06-17 2024-04-24 F. Hoffmann-La Roche AG Nucleoside-5 -oligophosphates having a cationically-modified nucleobase
US20250283162A1 (en) 2021-06-17 2025-09-11 Roche Sequencing Solutions, Inc. Engineered nanopore with a negatively charged polymer threaded through the channel
CN117999346A (zh) 2021-07-21 2024-05-07 豪夫迈·罗氏有限公司 形成窄通道孔的α-溶血素变体及其用途
WO2023089175A1 (en) 2021-11-22 2023-05-25 F. Hoffmann-La Roche Ag Thermostable polymerase variants
WO2026050292A1 (en) 2024-08-29 2026-03-05 Roche Sequencing Solutions, Inc. Optimized reagent introduction during a nanopore sequencing assay
WO2026050290A1 (en) 2024-08-29 2026-03-05 Roche Sequencing Solutions, Inc. Lipid composition and methods of use in a nanopore sequencing assay
WO2026050289A1 (en) 2024-08-29 2026-03-05 Roche Sequencing Solutions, Inc. Pre-and post-sequencing cleaning protocols for nanopore sequencing

Family Cites Families (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
CA2243985C (en) 1997-09-11 2004-08-10 F. Hoffmann-La Roche Ag Thermostable dna polymerases incorporating nucleoside triphosphates labeled with fluorescein family dyes
US7238485B2 (en) 2004-03-23 2007-07-03 President And Fellows Of Harvard College Methods and apparatus for characterizing polynucleotides
WO2006120433A1 (en) 2005-05-10 2006-11-16 Solexa Limited Improved polymerases
GB0509508D0 (en) 2005-05-10 2005-06-15 Solexa Ltd Improved polymerases
WO2009122191A1 (en) 2008-04-04 2009-10-08 It-Is International Ltd Thermal control system and method for chemical and biochemical reactions
CA2742594C (en) * 2008-11-03 2020-03-24 Kapa Biosystems, Inc. Modified type a dna polymerases
CA2995178C (en) 2010-02-08 2023-09-12 Genia Technologies, Inc. Method and system for forming a nanopore in a lipid bilayer
GB201002362D0 (en) 2010-02-11 2010-03-31 Isis Innovation Peptide tag systems that spontaneously form an irreversible link to protein partners via isopeptide bonds
US8936911B2 (en) 2010-09-22 2015-01-20 Pacific Biosciences Of California, Inc. Purified extended polymerase/template complex for sequencing
EP2652153B1 (en) 2010-12-17 2017-07-05 The Trustees of Columbia University in the City of New York Dna sequencing by synthesis using modified nucleotides and nanopore detection
EP2689028B1 (en) 2011-03-23 2017-08-30 Pacific Biosciences Of California, Inc. Isolation of polymerase-nucleic acid complexes and loading onto substrates
US10059984B2 (en) 2011-06-16 2018-08-28 The Regents Of The University Of California Salt-tolerant DNA polymerases
CN103857805B (zh) 2011-08-10 2017-07-18 生命技术公司 聚合酶组合物、制备和使用所述聚合酶组合物的方法
CA2876095A1 (en) 2012-06-15 2013-12-19 Genia Technologies, Inc. Chip set-up and high-accuracy nucleic acid sequencing
US9605309B2 (en) 2012-11-09 2017-03-28 Genia Technologies, Inc. Nucleic acid sequencing using tags
US9567630B2 (en) 2013-10-23 2017-02-14 Genia Technologies, Inc. Methods for forming lipid bilayers on biochips
JP6736565B2 (ja) 2015-02-02 2020-08-05 エフ．ホフマン−ラ ロシュ アーゲーＦ． Ｈｏｆｆｍａｎｎ−Ｌａ Ｒｏｃｈｅ Ａｋｔｉｅｎｇｅｓｅｌｌｓｃｈａｆｔ ポリメラーゼバリアント
JP2016171215A (ja) 2015-03-12 2016-09-23 株式会社東芝 半導体記憶装置
US10526588B2 (en) 2015-05-14 2020-01-07 Roche Sequencing Solutions, Inc. Polymerase variants and uses thereof
WO2017148862A1 (en) 2016-02-29 2017-09-08 Genia Technologies, Inc. Polymerase variants
CN108699539B (zh) 2016-02-29 2022-11-18 吉尼亚科技公司 外切核酸酶缺陷的聚合酶

Also Published As

Publication number Publication date
US10526588B2 (en) 2020-01-07
JP2018515103A (ja) 2018-06-14
US10968438B2 (en) 2021-04-06
US20160333327A1 (en) 2016-11-17
US11788073B2 (en) 2023-10-17
WO2016183403A3 (en) 2017-01-05
EP3294749A4 (en) 2018-10-31
CA2978194A1 (en) 2016-11-17
JP2021121221A (ja) 2021-08-26
CA3028797C (en) 2023-01-03
JP2019187424A (ja) 2019-10-31
EP3294749A2 (en) 2018-03-21
CN107636004A (zh) 2018-01-26
US20200087636A1 (en) 2020-03-19
CA3028797A1 (en) 2016-11-17
US20210189355A1 (en) 2021-06-24
WO2016183403A2 (en) 2016-11-17

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