Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells

被引：487

作者：

Kleinstiver, Benjamin P. ^{[1
,2
,3
,4
]}

Tsai, Shengdar Q. ^{[1
,2
,3
,4
]}

Prew, Michelle S. ^{[1
,2
,3
]}

Nguyen, Nhu T. ^{[1
,2
,3
]}

Welch, Moira M. ^{[1
,2
,3
]}

Lopez, Jose M. ^{[1
,2
,3
,4
]}

McCaw, Zachary R. ^{[1
,2
,5
]}

Aryee, Martin J. ^{[1
,2
,4
,5
]}

Joung, J. Keith ^{[1
,2
,3
,4
]}

机构：

[1] Massachusetts Gen Hosp, Mol Pathol Unit, Charlestown, MA 02129 USA

[2] Massachusetts Gen Hosp, Ctr Canc Res, Charlestown, MA 02129 USA

[3] Massachusetts Gen Hosp, Ctr Computat & Integrat Biol, Charlestown, MA 02129 USA

[4] Harvard Med Sch, Dept Pathol, Boston, MA 02115 USA

[5] Harvard TH Chan Sch Publ Hlth, Dept Biostat, Boston, MA USA

来源：

NATURE BIOTECHNOLOGY | 2016年 / 34卷 / 08期

基金：

加拿大自然科学与工程研究理事会; 美国国家卫生研究院;

关键词：

OFF-TARGET CLEAVAGE; RNA; SYSTEMS; ENDONUCLEASE; DELIVERY; TALENS; RANGE;

D O I：

10.1038/nbt.3620

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 activities and genome-wide specificities of CRISPR-Cas Cpf1 nucleases(1) are not well defined. We show that two Cpf1 nucleases from Acidaminococcus sp. BV3L6 and Lachnospiraceae bacterium ND2006 (AsCpf1 and LbCpf1, respectively) have on-target efficiencies in human cells comparable with those of the widely used Streptococcus pyogenes Cas9 (SpCas9)(2-5). We also report that four to six bases at the 3' end of the short CRISPR RNA (crRNA) used to program Cpf1 nucleases are insensitive to single base mismatches, but that many of the other bases in this region of the crRNA are highly sensitive to single or double substitutions. Using GUIDE-seq and targeted deep sequencing analyses performed with both Cpf1 nucleases, we were unable to detect off-target cleavage for more than half of 20 different crRNAs. Our results suggest that AsCpf1 and LbCpf1 are highly specific in human cells.

引用

页码：869 / +

页数：7

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