JP2024501832A - 高忠実度DNAポリメラーゼによるrhPCRベース増幅産物シーケンシングにおいて、プライマー二量体及びオフターゲット増幅を低減するRNアーゼH2突然変異体 - Google Patents

高忠実度DNAポリメラーゼによるrhPCRベース増幅産物シーケンシングにおいて、プライマー二量体及びオフターゲット増幅を低減するRNアーゼH2突然変異体 Download PDF

Info

Publication number
JP2024501832A
JP2024501832A JP2023538971A JP2023538971A JP2024501832A JP 2024501832 A JP2024501832 A JP 2024501832A JP 2023538971 A JP2023538971 A JP 2023538971A JP 2023538971 A JP2023538971 A JP 2023538971A JP 2024501832 A JP2024501832 A JP 2024501832A
Authority
JP
Japan
Prior art keywords
rnase
primer
dna polymerase
hybrid
sel29
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.)
Pending
Application number
JP2023538971A
Other languages
English (en)
Japanese (ja)
Other versions
JP2024501832A5 (https=
Inventor
ドボシー,ジョゼフ
フローリグ,ジョン
パーシュバチャー,キャサリン
ベルツ,クリスティン
ローズ,スコット
ベールケ,マーク・アーロン
Original Assignee
インテグレイテツド・デイー・エヌ・エイ・テクノロジーズ・インコーポレイテツド
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
Application filed by インテグレイテツド・デイー・エヌ・エイ・テクノロジーズ・インコーポレイテツド filed Critical インテグレイテツド・デイー・エヌ・エイ・テクノロジーズ・インコーポレイテツド
Publication of JP2024501832A publication Critical patent/JP2024501832A/ja
Publication of JP2024501832A5 publication Critical patent/JP2024501832A5/ja
Pending legal-status Critical Current

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844Nucleic acid amplification reactions
    • C12Q1/6848Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/16Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62DNA sequences coding for fusion proteins
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844Nucleic acid amplification reactions
    • C12Q1/6853Nucleic acid amplification reactions using modified primers or templates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y301/00Hydrolases acting on ester bonds (3.1)
    • C12Y301/26Endoribonucleases producing 5'-phosphomonoesters (3.1.26)
    • C12Y301/26004Ribonuclease H (3.1.26.4)
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/31Chemical structure of the backbone
    • C12N2310/315Phosphorothioates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/32Chemical structure of the sugar
    • C12N2310/3212'-O-R Modification
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/30Chemical structure
    • C12N2310/32Chemical structure of the sugar
    • C12N2310/3222'-R Modification
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844Nucleic acid amplification reactions
    • C12Q1/686Polymerase chain reaction [PCR]
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Plant Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Saccharide Compounds (AREA)
JP2023538971A 2020-12-24 2021-12-22 高忠実度DNAポリメラーゼによるrhPCRベース増幅産物シーケンシングにおいて、プライマー二量体及びオフターゲット増幅を低減するRNアーゼH2突然変異体 Pending JP2024501832A (ja)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US202063130548P 2020-12-24 2020-12-24
US63/130,548 2020-12-24
US202163277273P 2021-11-09 2021-11-09
US63/277,273 2021-11-09
PCT/US2021/064879 WO2022140553A1 (en) 2020-12-24 2021-12-22 Rnase h2 mutants that reduce primer dimers and off-target amplification in rhpcr-based amplicon sequencing with high-fidelity dna polymerases

Publications (2)

Publication Number Publication Date
JP2024501832A true JP2024501832A (ja) 2024-01-16
JP2024501832A5 JP2024501832A5 (https=) 2025-01-06

Family

ID=80050536

Family Applications (1)

Application Number Title Priority Date Filing Date
JP2023538971A Pending JP2024501832A (ja) 2020-12-24 2021-12-22 高忠実度DNAポリメラーゼによるrhPCRベース増幅産物シーケンシングにおいて、プライマー二量体及びオフターゲット増幅を低減するRNアーゼH2突然変異体

Country Status (8)

Country Link
US (2) US12024742B2 (https=)
EP (1) EP4267762B1 (https=)
JP (1) JP2024501832A (https=)
KR (1) KR20230124946A (https=)
AU (1) AU2021409723A1 (https=)
CA (1) CA3206360A1 (https=)
ES (1) ES3060434T3 (https=)
WO (1) WO2022140553A1 (https=)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011521624A (ja) * 2008-04-30 2011-07-28 インテグレイテツド・デイー・エヌ・エイ・テクノロジーズ・インコーポレイテツド 修飾されたrna単量体を用いるrnアーゼhを基礎とするアッセイ
WO2018031625A2 (en) * 2016-08-09 2018-02-15 Integrated Dna Technologies, Inc. Rnase h mutants in an emulsion
US20180057868A1 (en) * 2016-08-30 2018-03-01 Integrated Dna Technologies, Inc. Cleavable hairpin primers
WO2019215067A1 (en) * 2018-05-07 2019-11-14 Roche Innovation Center Copenhagen A/S Massively parallel discovery methods for oligonucleotide therapeutics
WO2020136440A2 (en) * 2018-12-28 2020-07-02 National University Of Singapore Methods for targeted complementary dna enrichment

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4458066A (en) 1980-02-29 1984-07-03 University Patents, Inc. Process for preparing polynucleotides
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
ATE138106T1 (de) 1988-07-20 1996-06-15 David Segev Verfahren zur amplifizierung und zum nachweis von nukleinsäuresequenzen
US5494810A (en) 1990-05-03 1996-02-27 Cornell Research Foundation, Inc. Thermostable ligase-mediated DNA amplifications system for the detection of genetic disease
DE60027040T2 (de) 1999-10-29 2006-11-23 Stratagene California, La Jolla Zusammensetzungen und methoden zur verwendung von dna polymerasen
US20090068643A1 (en) 2005-11-23 2009-03-12 Integrated Dna Technologies, Inc. Dual Function Primers for Amplifying DNA and Methods of Use
US8911948B2 (en) 2008-04-30 2014-12-16 Integrated Dna Technologies, Inc. RNase H-based assays utilizing modified RNA monomers
EP2333109B1 (en) * 2008-09-03 2013-08-07 Takara Bio, Inc. Composition for detection of rna
JP2016536995A (ja) * 2013-11-14 2016-12-01 インテグレイテツド・デイー・エヌ・エイ・テクノロジーズ・インコーポレイテツド 増大した鋳型識別活性を有するdnaポリメラーゼ突然変異体
HK1247255A1 (zh) 2014-12-15 2018-09-21 Cepheid 底数大於2的指数核酸扩增

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011521624A (ja) * 2008-04-30 2011-07-28 インテグレイテツド・デイー・エヌ・エイ・テクノロジーズ・インコーポレイテツド 修飾されたrna単量体を用いるrnアーゼhを基礎とするアッセイ
WO2018031625A2 (en) * 2016-08-09 2018-02-15 Integrated Dna Technologies, Inc. Rnase h mutants in an emulsion
US20180057868A1 (en) * 2016-08-30 2018-03-01 Integrated Dna Technologies, Inc. Cleavable hairpin primers
WO2019215067A1 (en) * 2018-05-07 2019-11-14 Roche Innovation Center Copenhagen A/S Massively parallel discovery methods for oligonucleotide therapeutics
WO2020136440A2 (en) * 2018-12-28 2020-07-02 National University Of Singapore Methods for targeted complementary dna enrichment

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
FRONT. MICROBIOL., vol. 11, no. 560791, JPN6025049145, 2020, pages 1 - 9, ISSN: 0005743931 *
JOURNAL OF BACTERIOLOGY, vol. 180, no. 23, JPN6025049144, 1998, pages 6207 - 6214, ISSN: 0005743930 *

Also Published As

Publication number Publication date
KR20230124946A (ko) 2023-08-28
WO2022140553A1 (en) 2022-06-30
AU2021409723A9 (en) 2024-09-12
US20220348997A1 (en) 2022-11-03
CA3206360A1 (en) 2022-06-30
EP4267762B1 (en) 2025-09-17
US20240318237A1 (en) 2024-09-26
AU2021409723A1 (en) 2023-06-22
EP4267762C0 (en) 2025-09-17
US12024742B2 (en) 2024-07-02
ES3060434T3 (en) 2026-03-26
EP4267762A1 (en) 2023-11-01

Similar Documents

Publication Publication Date Title
JP6510695B2 (ja) 修飾されたrna単量体を用いるrnアーゼhを基礎とするアッセイ
JP7640606B2 (ja) 増大した鋳型識別活性を有するdnaポリメラーゼ突然変異体
JP2016510601A (ja) 修飾RNAモノマーを用いたRNaseH系アッセイ
JP6316505B2 (ja) 易熱性エキソヌクレアーゼ
WO2012135053A2 (en) Rnase h-based assays utilizing modified rna monomers
WO2013142364A1 (en) Modified rnase h enzymes and their uses
US20170044507A1 (en) Modified rnase h enzymes and their uses
EP4267762B1 (en) Rnase h2 mutants that reduce primer dimers and off-target amplification in rhpcr-based amplicon sequencing with high-fidelity dna polymerases
CN116829720A (zh) 在使用高保真dna聚合酶的基于rhpcr的扩增子测序中减少引物二聚体和脱靶扩增的rna酶h2突变体

Legal Events

Date Code Title Description
A521 Request for written amendment filed

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20241220

A621 Written request for application examination

Free format text: JAPANESE INTERMEDIATE CODE: A621

Effective date: 20241220

A977 Report on retrieval

Free format text: JAPANESE INTERMEDIATE CODE: A971007

Effective date: 20251128

A131 Notification of reasons for refusal

Free format text: JAPANESE INTERMEDIATE CODE: A131

Effective date: 20251202

A601 Written request for extension of time

Free format text: JAPANESE INTERMEDIATE CODE: A601

Effective date: 20260226