	US 11,840,685 B2
	Inhibition of unintended mutations in gene editing
Jia Chen, Shanghai (CN); Bei Yang, Shanghai (CN); Li Yang, Shanghai (CN); Xingxu Huang, Shanghai (CN); and Lijie Wang, Shanghai (CN)
Assigned to ShanghaiTech University, Shanghai (CN)
Filed by ShanghaiTech University, Shanghai (CN)
Filed on Jul. 11, 2022, as Appl. No. 17/862,354.
Application 17/862,354 is a continuation of application No. 17/427,040, granted, now 11,384,353, previously published as PCT/CN2020/074218, filed on Feb. 3, 2020.
Claims priority of application No. PCT/CN2019/074577 (WO), filed on Feb. 2, 2019.
Prior Publication US 2023/0094769 A1, Mar. 30, 2023
Int. Cl. C12N 15/10 (2006.01); C12N 9/22 (2006.01); C12N 9/50 (2006.01); C12N 9/78 (2006.01); C12N 15/11 (2006.01)

CPC C12N 15/102 (2013.01) [C12N 9/22 (2013.01); C12N 9/506 (2013.01); C12N 9/78 (2013.01); C12N 15/111 (2013.01); C07K 2319/50 (2013.01); C07K 2319/85 (2013.01); C12N 2310/20 (2017.05); C12Y 304/22044 (2013.01); C12Y 305/04004 (2013.01); C12Y 305/04005 (2013.01)]

13 Claims

1. A dual guide RNA system, comprising:

a target single guide RNA comprising a first spacer having sequence complementarity to a target nucleic acid sequence proximate to a first protospacer adjacent motif (PAM) site,

a helper single guide RNA comprising a second spacer having sequence complementarity to a second nucleic acid sequence proximate to a second PAM site, wherein said second spacer is 8-15 bases in length,

a clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) protein, and

a nucleobase deaminase,

wherein the second PAM site is from 10 to 150 bases from the first PAM site.