WO2019014359A3 - Polymerase chain transcription (pct): exponential synthesis of rna and modified rna - Google Patents

Polymerase chain transcription (pct): exponential synthesis of rna and modified rna Download PDF

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Publication number
WO2019014359A3
WO2019014359A3 PCT/US2018/041660 US2018041660W WO2019014359A3 WO 2019014359 A3 WO2019014359 A3 WO 2019014359A3 US 2018041660 W US2018041660 W US 2018041660W WO 2019014359 A3 WO2019014359 A3 WO 2019014359A3
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WO
WIPO (PCT)
Prior art keywords
rna
generating
reaction mixture
pct
polymerase chain
Prior art date
Application number
PCT/US2018/041660
Other languages
French (fr)
Other versions
WO2019014359A2 (en
Inventor
Floyd Romesberg
Tingjian CHEN
Original Assignee
The Scripps Research Institute
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.)
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Publication date
Application filed by The Scripps Research Institute filed Critical The Scripps Research Institute
Publication of WO2019014359A2 publication Critical patent/WO2019014359A2/en
Publication of WO2019014359A3 publication Critical patent/WO2019014359A3/en

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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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/111—General methods applicable to biologically active non-coding nucleic acids
    • 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
    • C12N2310/10—Type of nucleic acid
    • C12N2310/16—Aptamers
    • 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
    • C12N2330/00—Production
    • C12N2330/30—Production chemically synthesised

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Provided herein are methods of generating a RNA, comprising: providing a reaction mixture comprising a DNA template, a forward primer, a reverse primer, a SFM4-3 polymerase, and ribonucleotides; and generating the RNA by thermocycling the reaction mixture. Also provided herein are methods for generating a RNA or modified RNA at an elevated temperature, the method comprising: contacting a reaction mixture comprising a DNA template, a primer, a SFM4-3 polymerase, and ribonucleoside triphosphates; and generating the RNA by incubation at the elevated temperature or putting the reaction mixture through repeated cycles of heating and cooling. Further provided herein are kits for transcribing, synthesizing, and/or amplifying a RNA sequence comprising: a polymerase; and ribonucleoside triphosphates.
PCT/US2018/041660 2017-07-12 2018-07-11 Polymerase chain transcription (pct): exponential synthesis of rna and modified rna WO2019014359A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201762531603P 2017-07-12 2017-07-12
US62/531,603 2017-07-12

Publications (2)

Publication Number Publication Date
WO2019014359A2 WO2019014359A2 (en) 2019-01-17
WO2019014359A3 true WO2019014359A3 (en) 2019-02-21

Family

ID=65001793

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2018/041660 WO2019014359A2 (en) 2017-07-12 2018-07-11 Polymerase chain transcription (pct): exponential synthesis of rna and modified rna

Country Status (1)

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WO (1) WO2019014359A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112813120A (en) * 2020-12-31 2021-05-18 华南理工大学 Method for efficiently synthesizing 5' labeled RNA by using DNA polymerase mutant strain

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5916777A (en) * 1995-06-07 1999-06-29 Gen-Probe Incorporated Enzymatic synthesis of oligonucleotides using 3'-ribonucleotide primers
US5939292A (en) * 1996-08-06 1999-08-17 Roche Molecular Systems, Inc. Thermostable DNA polymerases having reduced discrimination against ribo-NTPs
US20060003353A1 (en) * 2001-09-28 2006-01-05 Sigma-Aldrich Co. Recombinant DNA processes using a dNTP mixture containing modified nucleotides
WO2006039656A2 (en) * 2004-10-01 2006-04-13 Novartis Vaccines And Diagnostics Inc. Modified small interfering rna molecules and methods of use
US20060141467A1 (en) * 2002-12-20 2006-06-29 Q-Rna, Inc. Methods and kits for making double stranded ribonucleic acids
WO2007025195A1 (en) * 2005-08-24 2007-03-01 Datascope Investment Corporation Methods and kits for sense rna synthesis
US20070219148A1 (en) * 2003-07-02 2007-09-20 Commissariat A L'energie Atomique Small Interfering RNA Specific to Sub-Units $g(a),$g(a)' and $g(b) of the Kinase Protein ck2,and the Applications of the Same
US20110098461A1 (en) * 2000-08-02 2011-04-28 University Of Southern California Novel RNA Interference Methods Using DNA-RNA Duplex Constructs
US20140093938A1 (en) * 2009-07-21 2014-04-03 Biotec Pharmacon Asa Method of Removing Nucleic Acid Contamination in Reverse Transcription and Amplification Reactions

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5916777A (en) * 1995-06-07 1999-06-29 Gen-Probe Incorporated Enzymatic synthesis of oligonucleotides using 3'-ribonucleotide primers
US5939292A (en) * 1996-08-06 1999-08-17 Roche Molecular Systems, Inc. Thermostable DNA polymerases having reduced discrimination against ribo-NTPs
US20110098461A1 (en) * 2000-08-02 2011-04-28 University Of Southern California Novel RNA Interference Methods Using DNA-RNA Duplex Constructs
US20060003353A1 (en) * 2001-09-28 2006-01-05 Sigma-Aldrich Co. Recombinant DNA processes using a dNTP mixture containing modified nucleotides
US20060141467A1 (en) * 2002-12-20 2006-06-29 Q-Rna, Inc. Methods and kits for making double stranded ribonucleic acids
US20070219148A1 (en) * 2003-07-02 2007-09-20 Commissariat A L'energie Atomique Small Interfering RNA Specific to Sub-Units $g(a),$g(a)' and $g(b) of the Kinase Protein ck2,and the Applications of the Same
WO2006039656A2 (en) * 2004-10-01 2006-04-13 Novartis Vaccines And Diagnostics Inc. Modified small interfering rna molecules and methods of use
WO2007025195A1 (en) * 2005-08-24 2007-03-01 Datascope Investment Corporation Methods and kits for sense rna synthesis
US20140093938A1 (en) * 2009-07-21 2014-04-03 Biotec Pharmacon Asa Method of Removing Nucleic Acid Contamination in Reverse Transcription and Amplification Reactions

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CHEN ET AL.: "Evolution of Thermophilic DNA Polymerases for the Recognition and Amplification of C2'-Modified DNA", NATURE CHEMISTRY, vol. 8, no. 6, June 2016 (2016-06-01), pages 1 - 18, XP055571086 *
CHEN ET AL.: "Polymerase Chain Transcription: Exponential Synthesis of RNA and Modified RNA", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 139, no. 29, 26 July 2017 (2017-07-26), pages 9949 - 9954, XP055571087 *

Also Published As

Publication number Publication date
WO2019014359A2 (en) 2019-01-17

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