Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors

被引：344

作者：

Wang, Xiaoling ^{[1
]}

Wang, Yebo ^{[1
,2
]}

Wu, Xiwei ^{[3
]}

Wang, Jinhui ^{[3
]}

Wang, Yingjia ^{[1
,2
]}

Qiu, Zhaojun ^{[3
]}

Chang, Tammy ^{[1
]}

Huang, He ^{[2
]}

Lin, Ren-Jang ^{[3
]}

Yee, Jiing-Kuan ^{[1
]}

机构：

[1] Beckman Res Inst City Hope, Dept Virol, Duarte, CA 91010 USA

[2] Zhejiang Univ, Affiliated Hosp 1, Bone Marrow Transplantat Ctr, Hangzhou 310003, Zhejiang, Peoples R China

[3] Beckman Res Inst City Hope, Dept Mol & Cellular Biol, Duarte, CA USA

来源：

NATURE BIOTECHNOLOGY | 2015年 / 33卷 / 02期

基金：

中国国家自然科学基金;

关键词：

HUMAN-CELLS; NUCLEASE SPECIFICITY; CAS9; CRISPR/CAS; DNA; DELETIONS; NICKASES; SYSTEMS;

D O I：

10.1038/nbt.3127

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The utility of CRISPR-Cas9 and TALENs for genome editing may be compromised by their off-target activity. We show that integrase-defective lentiviral vectors (IDLVs) can detect such off-target cleavage with a frequency as low as 1%. In the case of Cas9, we find frequent off-target sites with a one-base bulge or up to 13 mismatches between the single guide RNA (sgRNA) and its genomic target; which refines sgRNA design.

引用

页码：175 / 178

页数：4

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