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1. WO2020109412 - TARGETED ENRICHMENT BY ENDONUCLEASE PROTECTION

Publication Number WO/2020/109412
Publication Date 04.06.2020
International Application No. PCT/EP2019/082791
International Filing Date 27.11.2019
IPC
C12Q 1/68 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
C12Q 1/6806 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
6806Preparing nucleic acids for analysis, e.g. for polymerase chain reaction assay
CPC
C12Q 1/6806
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
6806Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
Applicants
  • KEYGENE N.V. [NL]/[NL]
Inventors
  • WHITE, Stefan John
  • HOGERS, René Cornelis Josephus
Agents
  • NEDERLANDSCH OCTROOIBUREAU
Priority Data
18208936.728.11.2018EP
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) TARGETED ENRICHMENT BY ENDONUCLEASE PROTECTION
(FR) ENRICHISSEMENT CIBLÉ PAR PROTECTION D'ENDONUCLÉASE
Abstract
(EN)
The current invention pertains to a method for the enrichment of a target nucleic acid fragment from a nucleic acid sample, comprising the steps of cleaving the nucleic acid sample with a first and a second RNA guided or DNA guided endonuclease complex, preferably a first and a second gRNA-CAS complex, thereby generating the target nucleic acid fragment and at least one non-target nucleic acid fragment. The generated fragments are subsequently contacted with an exonuclease, wherein the exonuclease digests only the non-target nucleic acid fragments. The invention further pertains to the use of the enriched target nucleic acid fragments for preparing an adapter ligated target nucleic acid fragment and for sequencing the target nucleic acid fragment.
(FR)
La présente invention concerne un procédé d'enrichissement d'un fragment d'acide nucléique cible à partir d'un échantillon d'acide nucléique, consistant à procéder au clivage de l'échantillon d'acide nucléique avec un premier et un second complexe d'endonucléase guidé par ARN ou ADN, de préférence un premier et un second complexe ARNg-CAS, produisant ainsi le fragment d'acide nucléique cible et au moins un fragment d'acide nucléique non cible. Les fragments produits sont ensuite mis en contact avec une exonucléase digérant uniquement les fragments d'acide nucléique non cibles. L'invention concerne en outre l'utilisation des fragments d'acide nucléique cibles enrichis pour préparer un fragment d'acide nucléique cible ligaturé par un adaptateur et pour séquencer le fragment d'acide nucléique cible.
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