Title:
METHOD FOR IMPROVING DETECTION ACCURACY OF LOW-FREQUENCY SINGLE NUCLEOTIDE VARIANTS BY USING CRISPR/CAS9 SYSTEM IN NEXT-GENERATION SEQUENCING
Document Type and Number:
WIPO Patent Application WO/2022/086153
Kind Code:
A1
Abstract:
The present invention relates to a method for detecting single nucleotide variants by using CRISPR/Cas9 system. The present invention can more accurately detect single nucleotide variants by excluding false positive SNV detection caused by technical errors during next-generation sequencing (NGS), and accurately targets and labels desired SNVs by using CRISPR/Cas9 system, and thus can accurately detect low-frequency SNVs.
Inventors:
BANG DU HEE (KR)
LEE DONG IN (KR)
LEE DONG IN (KR)
Application Number:
PCT/KR2021/014666
Publication Date:
April 28, 2022
Filing Date:
October 20, 2021
Export Citation:
Assignee:
UNIV YONSEI IACF (KR)
International Classes:
C12Q1/6886; C12N9/22; C12N15/10; C12N15/113
Foreign References:
| CN108342480A | 2018-07-31 |
Other References:
HWANG GUE-HO, YU JIHYEON, YANG SOYEON, SON WOO JAE, LIM KAYEONG, KIM HEON SEOK, KIM JIN-SOO, BAE SANGSU: "CRISPR-sub: Analysis of DNA substitution mutations caused by CRISPR-Cas9 in human cells", COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL, RESEARCH NETWORK OF COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY, SWEDEN, vol. 18, 1 January 2020 (2020-01-01), Sweden , pages 1686 - 1694, XP055923680, ISSN: 2001-0370, DOI: 10.1016/j.csbj.2020.06.026
KANG SEUNG-HUN, LEE WI-JAE, AN JU-HYUN, LEE JONG-HEE, KIM YOUNG-HYUN, KIM HANSEOP, OH YEOUNSUN, PARK YOUNG-HO, JIN YEUNG BAE, JUN : "Prediction-based highly sensitive CRISPR off-target validation using target-specific DNA enrichment", NATURE COMMUNICATIONS, vol. 11, no. 1, 1 December 2020 (2020-12-01), XP055923681, DOI: 10.1038/s41467-020-17418-8
SINGH RAJESH R.: "Next-Generation Sequencing in High-Sensitive Detection of Mutations in Tumors", THE JOURNAL OF MOLECULAR DIAGNOSTICS, AMERICAN SOCIETY FOR INVESTIGATIVE PATHOLOGY AND THE ASSOCIATION FOR MOLECULAR PATHOLOGY, vol. 22, no. 8, 1 August 2020 (2020-08-01), pages 994 - 1007, XP055923682, ISSN: 1525-1578, DOI: 10.1016/j.jmoldx.2020.04.213
KIM JUNHO, KIM DACHAN, LIM JAE SEOK, MAENG JU HEON, SON HYEONJU, KANG HOON-CHUL, NAM HOJUNG, LEE JEONG HO, KIM SANGWOO: "The use of technical replication for detection of low-level somatic mutations in next-generation sequencing", NATURE COMMUNICATIONS, vol. 10, no. 1, 1 December 2019 (2019-12-01), XP055780865, DOI: 10.1038/s41467-019-09026-y
LEE DONGIN, LEE JI HYUN, BANG DUHEE: "Accurate Detection of Rare Mutant Alleles by Target Base-Specific Cleavage with the CRISPR/Cas9 System", ACS SYNTHETIC BIOLOGY, AMERICAN CHEMICAL SOCIETY, WASHINGTON DC ,USA, vol. 10, no. 6, 18 June 2021 (2021-06-18), Washington DC ,USA , pages 1451 - 1464, XP055923683, ISSN: 2161-5063, DOI: 10.1021/acssynbio.1c00056
KANG SEUNG-HUN, LEE WI-JAE, AN JU-HYUN, LEE JONG-HEE, KIM YOUNG-HYUN, KIM HANSEOP, OH YEOUNSUN, PARK YOUNG-HO, JIN YEUNG BAE, JUN : "Prediction-based highly sensitive CRISPR off-target validation using target-specific DNA enrichment", NATURE COMMUNICATIONS, vol. 11, no. 1, 1 December 2020 (2020-12-01), XP055923681, DOI: 10.1038/s41467-020-17418-8
SINGH RAJESH R.: "Next-Generation Sequencing in High-Sensitive Detection of Mutations in Tumors", THE JOURNAL OF MOLECULAR DIAGNOSTICS, AMERICAN SOCIETY FOR INVESTIGATIVE PATHOLOGY AND THE ASSOCIATION FOR MOLECULAR PATHOLOGY, vol. 22, no. 8, 1 August 2020 (2020-08-01), pages 994 - 1007, XP055923682, ISSN: 1525-1578, DOI: 10.1016/j.jmoldx.2020.04.213
KIM JUNHO, KIM DACHAN, LIM JAE SEOK, MAENG JU HEON, SON HYEONJU, KANG HOON-CHUL, NAM HOJUNG, LEE JEONG HO, KIM SANGWOO: "The use of technical replication for detection of low-level somatic mutations in next-generation sequencing", NATURE COMMUNICATIONS, vol. 10, no. 1, 1 December 2019 (2019-12-01), XP055780865, DOI: 10.1038/s41467-019-09026-y
LEE DONGIN, LEE JI HYUN, BANG DUHEE: "Accurate Detection of Rare Mutant Alleles by Target Base-Specific Cleavage with the CRISPR/Cas9 System", ACS SYNTHETIC BIOLOGY, AMERICAN CHEMICAL SOCIETY, WASHINGTON DC ,USA, vol. 10, no. 6, 18 June 2021 (2021-06-18), Washington DC ,USA , pages 1451 - 1464, XP055923683, ISSN: 2161-5063, DOI: 10.1021/acssynbio.1c00056
Attorney, Agent or Firm:
DANA PATENT LAW FIRM (KR)
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