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1. WO2021058145 - PHAGE T7 PROMOTERS FOR BOOSTING IN VITRO TRANSCRIPTION

Publication Number WO/2021/058145
Publication Date 01.04.2021
International Application No. PCT/EP2020/066070
International Filing Date 10.06.2020
IPC
C12N 15/70 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70Vectors or expression systems specially adapted for E. coli
C12Q 1/6865 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6844Nucleic acid amplification reactions
6865Promoter-based amplification, e.g. nucleic acid sequence-based amplification , self-sustained sequence replication or transcription-based amplification system
C12N 15/10 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
CPC
C12N 15/1096
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
1096cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
C12Q 1/6865
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6844Nucleic acid amplification reactions
6865Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS]
Applicants
  • MAX-DELBRÜCK-CENTRUM FÜR MOLEKULARE MEDIZIN IN DER HELMHOLTZ-GEMEINSCHAFT [DE]/[DE]
Inventors
  • CONRAD, Thomas
  • SAUER, Sascha
Agents
  • VOSSIUS & PARTNER (NO 31)
Priority Data
19199289.024.09.2019EP
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) PHAGE T7 PROMOTERS FOR BOOSTING IN VITRO TRANSCRIPTION
(FR) PROMOTEURS DE PHAGE T7 POUR AMPLIFIER LA TRANSCRIPTION IN VITRO
Abstract
(EN)
The invention relates to a DNA polynucleotide, a kit comprising said DNA polynucleotide, and a method for in vitro transcribing (IVT) RNA using said DNA polynucleotide, wherein said DNA polynucleotide comprises (i) a T7 promoter sequence comprising (1) a T7 core promoter sequence and (2) a directly adjacent downstream flanking region, wherein the downstream flanking region comprises eight bases (+1 to +8) comprising three guanines at positions +1 to +3, an adenine or thymine at position +4, at least two adenines at positions +4 to +8 and/or a thymine at position +4, and at most one cytosine at positions +4 to +8, and downstream thereof (ii) a nucleotide sequence encoding the RNA to be transcribed; and wherein the method comprises (a) providing said DNA polynucleotide as an IVT template, and (b) in vitro transcribing said IVT template in the presence of a T7 polymerase and ribonucleotide triphosphates.
(FR)
L'invention concerne un polynucléotide d'ADN, un kit comprenant ledit polynucléotide d'ADN, et un procédé de transcription in vitro d'ARN À l'aide dudit polynucléotide d'ADN, ledit polynucléotide d'ADN comprenant (i) une séquence de promoteur T7 comprenant (1) une séquence de promoteur principal T7 et (2) une région de flanquement en aval directement adjacente, la région de flanquement en aval comprenant huit bases (+1 à +8) comprenant trois guanines à des positions +1 à +3, une adénine ou une thymine en position +4, au moins deux adénosines à des positions +4 à +8 et/ou une thymine en position +4, et au plus une cytosine à des positions +4 à +8, et en aval de celle-ci (ii) une séquence nucléotidique codant pour l'ARN à transcrire; et le procédé comprenant (a) la fourniture dudit polynucléotide d'ADN en tant que matrice IVT, et (b) à transcrire in vitro ledit modèle IVT en présence d'une polymérase T7 et de ribonucléotides triphosphates.
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