CA3028797C - Polymerase variants and uses thereof - Google Patents

Polymerase variants and uses thereof Download PDF

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Publication number
CA3028797C
CA3028797C CA3028797A CA3028797A CA3028797C CA 3028797 C CA3028797 C CA 3028797C CA 3028797 A CA3028797 A CA 3028797A CA 3028797 A CA3028797 A CA 3028797A CA 3028797 C CA3028797 C CA 3028797C
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CA
Canada
Prior art keywords
polymerase
dna
nucleotide
amino acid
nanopore
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
CA3028797A
Other languages
English (en)
French (fr)
Other versions
CA3028797A1 (en
Inventor
Aruna Ayer
Cleoma Arnold
Charles Schwab
Ilya Lederman
Colin Mcgaw
Tyler Shultz
Eileen Thai
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
Publication of CA3028797A1 publication Critical patent/CA3028797A1/en
Application granted granted Critical
Publication of CA3028797C publication Critical patent/CA3028797C/en
Active legal-status Critical Current
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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

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
CA3028797A 2015-05-14 2016-05-13 Polymerase variants and uses thereof Active CA3028797C (en)

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 2016-02-01
US15/012,317 US10308918B2 (en) 2015-02-02 2016-02-01 Polymerase variants
US15/151,364 US10526588B2 (en) 2015-05-14 2016-05-10 Polymerase variants and uses thereof
US15/151,364 2016-05-10
CA2978194A CA2978194C (en) 2015-05-14 2016-05-13 Polymerase variants and uses thereof

Related Parent Applications (1)

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

Publications (2)

Publication Number Publication Date
CA3028797A1 CA3028797A1 (en) 2016-11-17
CA3028797C true CA3028797C (en) 2023-01-03

Family

ID=57248698

Family Applications (2)

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

Family Applications After (1)

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

Country Status (6)

Country Link
US (3) US10526588B2 (2)
EP (1) EP3294749A4 (2)
JP (3) JP2018515103A (2)
CN (1) CN107636004A (2)
CA (2) CA3028797C (2)
WO (1) WO2016183403A2 (2)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016124543A1 (en) * 2015-02-02 2016-08-11 F. Hoffmann-La Roche Ag Polymerase variants
US10526588B2 (en) * 2015-05-14 2020-01-07 Roche Sequencing Solutions, Inc. Polymerase variants and uses thereof
EP3377648B1 (en) 2015-11-16 2021-01-06 Stratos Genomics, Inc. Dp04 polymerase variants
US10590480B2 (en) 2016-02-29 2020-03-17 Roche Sequencing Solutions, Inc. Polymerase variants
US20170268052A1 (en) 2016-02-29 2017-09-21 Genia Technologies, Inc. Polymerase-template complexes
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
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
JP7157048B2 (ja) 2016-09-22 2022-10-19 エフ．ホフマン－ラ ロシュ アーゲー Ｐｏｌ６ポリメラーゼバリアント
US11708566B2 (en) 2017-05-04 2023-07-25 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
WO2019166458A1 (en) 2018-02-28 2019-09-06 F. Hoffmann-La Roche Ag Alpha-hemolysin variants and uses thereof
CN115052882B (zh) 2020-02-06 2024-05-24 豪夫迈·罗氏有限公司 减少模板穿入纳米孔的组合物
ES2999861T3 (en) 2020-08-11 2025-02-27 Hoffmann La Roche Nucleoside-5'-oligophosphates tagged with positively-charged polymers, nanopores incorporating negative charges, and methods and systems using the same
WO2022263496A1 (en) 2021-06-17 2022-12-22 F. Hoffmann-La Roche Ag Engineered nanopore with a negatively charged polymer threaded through the channel
WO2022263489A1 (en) 2021-06-17 2022-12-22 F. Hoffmann-La Roche Ag Nucleoside-5 -oligophosphates having a cationically-modified nucleobase
EP4373925A1 (en) 2021-07-21 2024-05-29 F. Hoffmann-La Roche AG Alpha-hemolysin variants forming narrow channel pores and uses thereof
EP4437090A1 (en) 2021-11-22 2024-10-02 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)

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Publication number Priority date Publication date Assignee Title
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US6346379B1 (en) * 1997-09-11 2002-02-12 F. Hoffman-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
GB0509508D0 (en) 2005-05-10 2005-06-15 Solexa Ltd Improved polymerases
EP1888743B1 (en) 2005-05-10 2011-08-03 Illumina Cambridge Limited Improved polymerases
US9266109B2 (en) 2008-04-04 2016-02-23 It-Is International Ltd. Thermal control system and method for chemical and biochemical reactions
WO2010062777A2 (en) 2008-11-03 2010-06-03 Kapabiosystems Modified type a dna polymerases
WO2011097028A1 (en) 2010-02-08 2011-08-11 Genia Technologies, Inc. Systems and methods for manipulating a molecule in a nanopore
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
WO2012083249A2 (en) 2010-12-17 2012-06-21 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
WO2012173905A1 (en) 2011-06-16 2012-12-20 The Regents Of The Unversity Of California Salt-tolerant dna polymerases
US20130040365A1 (en) 2011-08-10 2013-02-14 Life Technologies Corporation Polymerase compositions, methods of making and using same
WO2013188841A1 (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
US9322062B2 (en) 2013-10-23 2016-04-26 Genia Technologies, Inc. Process for biosensor well formation
WO2016124543A1 (en) 2015-02-02 2016-08-11 F. Hoffmann-La Roche Ag Polymerase variants
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
ES2880331T3 (es) 2016-02-29 2021-11-24 Genia Tech Inc Variantes de polimerasa
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
US11788073B2 (en) 2023-10-17
JP2019187424A (ja) 2019-10-31
WO2016183403A2 (en) 2016-11-17
CA2978194A1 (en) 2016-11-17
EP3294749A2 (en) 2018-03-21
US20160333327A1 (en) 2016-11-17
CN107636004A (zh) 2018-01-26
US20200087636A1 (en) 2020-03-19
US10526588B2 (en) 2020-01-07
US10968438B2 (en) 2021-04-06
CA2978194C (en) 2021-11-16
US20210189355A1 (en) 2021-06-24
WO2016183403A3 (en) 2017-01-05
JP2018515103A (ja) 2018-06-14
JP2021121221A (ja) 2021-08-26
EP3294749A4 (en) 2018-10-31
CA3028797A1 (en) 2016-11-17

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