A highly specific CRISPR-Cas12j nuclease enables allele- specific genome editing

被引：38

作者：

Wang, Yao ^{[1
]}

Qi, Tao ^{[1
]}

Liu, Jingtong ^{[1
]}

Yang, Yuan ^{[1
]}

Wang, Ziwen ^{[2
]}

Wang, Ying ^{[3
]}

Wang, Tianyi ^{[1
]}

Li, Miaomiao ^{[1
]}

Li, Mingqing ^{[4
]}

Lu, Daru ^{[1
,5
]}

Chang, Alex Chia Yu ^{[6
,7
]}

Yang, Li ^{[8
,9
]}

Gao, Song ^{[2
]}

Wang, Yongming ^{[1
,10
]}

Lan, Feng ^{[1
,11
]}

机构：

[1] Fudan Univ, Zhongshan Hosp, Sch Life Sci, State Key Lab Genet Engn, Shanghai 200438, Peoples R China

[2] Sun Yat Sen Univ, Collaborat Innovat Ctr Canc Med, Canc Ctr, State Key Lab Oncol South China, Guangzhou 510060, Peoples R China

[3] Chinese Acad Sci, Univ Chinese Acad Sci, Shanghai Inst Nutr & Hlth, CAS Key Lab Computat Biol, Shanghai 200031, Peoples R China

[4] Fudan Univ, Hosp Obstet & Gynecol, Shanghai Inst Planned Parenthood Res, Shanghai Med Sch,Lab Reprod Immunol,NHC Key Lab Re, Shanghai 200080, Peoples R China

[5] Chongqing Populat & Family Planning Sci & Technol, NHC Key Lab Birth Defects & Reprod Hlth, Chongqing Key Lab Birth Defects & Reprod Hlth, Chongqing 400020, Peoples R China

[6] Shanghai Jiao Tong Univ, Peoples Hosp 9, Sch Med, Dept Cardiol, Shanghai 200125, Peoples R China

[7] Shanghai Jiao Tong Univ, Peoples Hosp 9, Shanghai Inst Precis Med, Sch Med, Shanghai 200125, Peoples R China

[8] Fudan Univ, Childrens Hosp, Ctr Mol Med, Shanghai 201102, Peoples R China

[9] Fudan Univ, Inst Biomed Sci, Shanghai Key Lab Med Epigenet, Int Lab Med Epigenet & Metab,Minist Sci & Technol, Shanghai 201102, Peoples R China

[10] Shanghai Engn Res Ctr Ind Microorganisms, Shanghai 200438, Peoples R China

[11] Chinese Acad Med Sci & Peking Union Med Coll, Fuwai Hosp, Natl Ctr Cardiovasc Dis, State Key Lab Cardiovasc Dis, Beijing 100037, Peoples R China

来源：

SCIENCE ADVANCES | 2023年 / 9卷 / 06期

基金：

中国国家自然科学基金;

关键词：

DNA; BASE; ENDONUCLEASE; MUTATION;

D O I：

10.1126/sciadv.abo6405

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The CRISPR-Cas system can treat autosomal dominant diseases by nonhomologous end joining (NHEJ) gene disruption of mutant alleles. However, many single-nucleotide mutations cannot be discriminated from wild -type alleles by current CRISPR-Cas systems. Here, we functionally screened six Cas12j nucleases and determined Cas12j-8 as an ideal genome editor with a hypercompact size. Cas12j-8 displayed comparable activity to AsCas12a and Un1Cas12f1. Cas12j-8 is a highly specific nuclease sensitive to single-nucleotide mismatches in the protospacer adjacent motif (PAM)-proximal region. We experimentally proved that Cas12j-8 enabled allele -specific disruption of genes with a single-nucleotide polymorphism (SNP). Cas12j-8 recognizes a simple TTN PAM that provides for high target site density. In silico analysis reveals that Cas12j-8 enables allele-specific dis-ruption of 25,931 clinically relevant variants in the ClinVar database, and 485,130,147 SNPs in the dbSNP data-base. Therefore, Cas12j-8 would be particularly suitable for therapeutic applications.

引用

页数：11

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