US 12,215,315 B2
Method for modifying target nucleic acid in genome of cell
Hyong Bum Kim, Seoul (KR); and Hui Kwon Kim, Jincheon-gun (KR)
Assigned to INDUSTRY-ACADEMIC COOPERATION FOUNDATION, YONSEI UNIVERSITY, Seoul (KR)
Appl. No. 17/271,971
Filed by INDUSTRY-ACADEMIC COOPERATION FOUNDATION, YONSEI UNIVERSITY, Seoul (KR)
PCT Filed Aug. 30, 2019, PCT No. PCT/KR2019/011166
§ 371(c)(1), (2) Date Feb. 26, 2021,
PCT Pub. No. WO2020/046047, PCT Pub. Date Mar. 5, 2020.
Claims priority of application No. 10-2018-0103979 (KR), filed on Aug. 31, 2018; and application No. 10-2019-0000391 (KR), filed on Jan. 2, 2019.
Prior Publication US 2021/0340528 A1, Nov. 4, 2021
Int. Cl. C12N 15/11 (2006.01); C12N 9/22 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/11 (2013.01) [C12N 9/22 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 4 Claims
OG exemplary drawing
 
1. A method for modifying a target nucleic acid in the genome of a cell, the method comprising:
incubating a cell comprising a target nucleic acid; a polynucleotide encoding a clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nuclease or a variant thereof, and a guide RNA,
wherein the Cas nuclease or a variant thereof is Streptococcus pyogenes Cas9-NG (SpCas9-NG),
wherein the target nucleic acid comprises a protospacer adjacent motif (PAM) and a target sequence complementary to the guide RNA,
the PAM consists of a sequence selected from the group consisting of 5′-NTG-3′, 5′-GTH-3′, 5′-VCGD-3′, and 5-TCGDG-3′,
the target nucleic acid can be recognized by a complex of the Cas nuclease or a variant thereof with the guide RNA, and
the complex of the Cas nuclease or a variant thereof with the guide RNA modifies the target nucleic acid sequence-specifically.