Title:
METHOD FOR GENERATING TOTAL MRNA-BASED RANDOM SGRNA LIBRARY THROUGH ENZYMATIC REACTION
Document Type and Number:
WIPO Patent Application WO/2024/072006
Kind Code:
A1
Abstract:
The present invention relates to a method for generating a total mRNA-based random sgRNA library through an enzymatic reaction. The method according to the present invention may generate an sgRNA library that may target the entire genome of any organism, including prokaryotes, without relying on genome sequence information and may be used for phenotype-based screening in prokaryotes because the generated sgRNAs selectively bind only to non-template strands of a target gene. In addition, mRNA, which is a useful genetic material, as well as DNA may be used as a starting material, enabling the generation of an sgRNA library targeting an actively expressed gene.
Inventors:
LEE JEONG WOOK (KR)
LEE JISEON (KR)
CHO BYUNG KWAN (KR)
JEON HAHYEON (KR)
CHOE DONG HUI (KR)
LEE JISEON (KR)
CHO BYUNG KWAN (KR)
JEON HAHYEON (KR)
CHOE DONG HUI (KR)
Application Number:
PCT/KR2023/014843
Publication Date:
April 04, 2024
Filing Date:
September 26, 2023
Export Citation:
Assignee:
POSTECH RES & BUSINESS DEV FOUND (KR)
KOREA ADVANCED INST SCI & TECH (KR)
KOREA ADVANCED INST SCI & TECH (KR)
International Classes:
C12N15/10; C12N15/113; C12Q1/6806; C40B50/06
Foreign References:
| KR20170020091A | 2017-02-22 | |||
| US20180051278A1 | 2018-02-22 |
Other References:
ARAKAWA HIROSHI: "A method to convert mRNA into a gRNA library for CRISPR/Cas9 editing of any organism", SCIENCE ADVANCES, vol. 2, no. 8, 5 August 2016 (2016-08-05), pages 1 - 10, XP055909688, DOI: 10.1126/sciadv.1600699
CRAVERO KAREN, MEDFORD ARIELLE, PALLAVAJJALA APARNA, CANZONIERO JENNA, HUNTER NATASHA, CHU DAVID, COCHRAN RORY L., WATERS IAN, CHR: "Biotinylated amplicon sequencing: A method for preserving DNA samples of limited quantity", PRACTICAL LABORATORY MEDICINE, vol. 12, 1 November 2018 (2018-11-01), pages 1 - 10, XP093151550, ISSN: 2352-5517, DOI: 10.1016/j.plabm.2018.e00108
BALDWIN AMBER, MORRIS ADAM R., MUKHERJEE NEELANJAN: "An Easy, Cost‐Effective, and Scalable Method to Deplete Human Ribosomal RNA for RNA‐seq", CURRENT PROTOCOLS, vol. 1, no. 6, 1 June 2021 (2021-06-01), pages 1 - 13, XP093151555, ISSN: 2691-1299, DOI: 10.1002/cpz1.176
CRAVERO KAREN, MEDFORD ARIELLE, PALLAVAJJALA APARNA, CANZONIERO JENNA, HUNTER NATASHA, CHU DAVID, COCHRAN RORY L., WATERS IAN, CHR: "Biotinylated amplicon sequencing: A method for preserving DNA samples of limited quantity", PRACTICAL LABORATORY MEDICINE, vol. 12, 1 November 2018 (2018-11-01), pages 1 - 10, XP093151550, ISSN: 2352-5517, DOI: 10.1016/j.plabm.2018.e00108
BALDWIN AMBER, MORRIS ADAM R., MUKHERJEE NEELANJAN: "An Easy, Cost‐Effective, and Scalable Method to Deplete Human Ribosomal RNA for RNA‐seq", CURRENT PROTOCOLS, vol. 1, no. 6, 1 June 2021 (2021-06-01), pages 1 - 13, XP093151555, ISSN: 2691-1299, DOI: 10.1002/cpz1.176
Attorney, Agent or Firm:
KIM, Kyeongkyo et al. (KR)
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