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

共 15 条

[11]

Perez-Pinera P, 2013, NAT METHODS, V10, P973, DOI [10.1038/NMETH.2600, 10.1038/nmeth.2600]

[12] Double Nicking by RNA-Guided CRISPR Cas9 for Enhanced Genome Editing Specificity [J].

Ran, F. Ann ;

Hsu, Patrick D. ;

Lin, Chie-Yu ;

Gootenberg, Jonathan S. ;

Konermann, Silvana ;

Trevino, Alexandro E. ;

Scott, David A. ;

Inoue, Azusa ;

Matoba, Shogo ;

Zhang, Yi ;

Zhang, Feng .

CELL, 2013, 154 (06) :1380-1389

[13] Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells [J].

Shalem, Ophir ;

Sanjana, Neville E. ;

Hartenian, Ella ;

Shi, Xi ;

Scott, David A. ;

Mikkelsen, Tarjei S. ;

Heckl, Dirk ;

Ebert, Benjamin L. ;

Root, David E. ;

Doench, John G. ;

Zhang, Feng .

SCIENCE, 2014, 343 (6166) :84-87

[14] Genetic Screens in Human Cells Using the CRISPR-Cas9 System [J].

Wang, Tim ;

Wei, Jenny J. ;

Sabatini, David M. ;

Lander, Eric S. .

SCIENCE, 2014, 343 (6166) :80-84

[15] Chromosomal deletions and inversions mediated by TALENs and CRISPR/Cas in zebrafish [J].

Xiao, An ;

Wang, Zhanxiang ;

Hu, Yingying ;

Wu, Yingdan ;

Luo, Zhou ;

Yang, Zhipeng ;

Zu, Yao ;

Li, Wenyuan ;

Huang, Peng ;

Tong, Xiangjun ;

Zhu, Zuoyan ;

Lin, Shuo ;

Zhang, Bo .

NUCLEIC ACIDS RESEARCH, 2013, 41 (14) :e141

← 1 2 →