CA3206360A1 - Rnase h2 mutants that reduce primer dimers and off-target amplification in rhpcr-based amplicon sequencing with high-fidelity dna polymerases - Google Patents

Rnase h2 mutants that reduce primer dimers and off-target amplification in rhpcr-based amplicon sequencing with high-fidelity dna polymerases

Info

Publication number
CA3206360A1
CA3206360A1 CA3206360A CA3206360A CA3206360A1 CA 3206360 A1 CA3206360 A1 CA 3206360A1 CA 3206360 A CA3206360 A CA 3206360A CA 3206360 A CA3206360 A CA 3206360A CA 3206360 A1 CA3206360 A1 CA 3206360A1
Authority
CA
Canada
Prior art keywords
rnase
primer
dna polymerase
hybrid
protein
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
CA3206360A
Other languages
English (en)
French (fr)
Inventor
Joseph Dobosy
John FROEHLIG
Katherine PERSCHBACHER
Kristin BELTZ
Scott Rose
Mark Aaron Behlke
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.)
Integrated DNA Technologies Inc
Original Assignee
Integrated DNA Technologies 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
Application filed by Integrated DNA Technologies Inc filed Critical Integrated DNA Technologies Inc
Publication of CA3206360A1 publication Critical patent/CA3206360A1/en
Pending legal-status Critical Current

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    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • 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
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/26—Endoribonucleases producing 5'-phosphomonoesters (3.1.26)
    • C12Y301/26004—Ribonuclease H (3.1.26.4)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/315—Phosphorothioates
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/321—2'-O-R Modification
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/32—Chemical structure of the sugar
    • C12N2310/322—2'-R Modification
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

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  • 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)
CA3206360A 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 Pending CA3206360A1 (en)

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 (1)

Publication Number Publication Date
CA3206360A1 true CA3206360A1 (en) 2022-06-30

Family

ID=80050536

Family Applications (1)

Application Number Title Priority Date Filing Date
CA3206360A Pending CA3206360A1 (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

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=)

Family Cites Families (16)

* 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
EP2644708B1 (en) * 2008-04-30 2014-12-17 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
WO2018031625A2 (en) * 2016-08-09 2018-02-15 Integrated Dna Technologies, Inc. Rnase h mutants in an emulsion
WO2018044831A1 (en) * 2016-08-30 2018-03-08 Integrated Dna Technologies, Inc. Cleavable hairpin primers
CN112105745A (zh) * 2018-05-07 2020-12-18 罗氏创新中心哥本哈根有限公司 用于寡核苷酸治疗剂的大规模平行发现方法
JP2022515466A (ja) * 2018-12-28 2022-02-18 ナショナル ユニヴァーシティー オブ シンガポール 標的化された相補的ｄｎａ富化のための方法

Also Published As

Publication number Publication date
KR20230124946A (ko) 2023-08-28
WO2022140553A1 (en) 2022-06-30
AU2021409723A9 (en) 2024-09-12
JP2024501832A (ja) 2024-01-16
US20220348997A1 (en) 2022-11-03
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

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