Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease

被引：200

作者：

Dahlman, James E. ^{[1
,2
]}

Abudayyeh, Omar O. ^{[1
,3
,4
,5
]}

Joung, Julia ^{[1
,3
,4
,5
]}

Gootenberg, Jonathan S. ^{[1
,3
,4
,5
,6
]}

Zhang, Feng ^{[1
,3
,4
,5
]}

Konermann, Silvana ^{[1
,3
,4
,5
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[2] Georgia Inst Technol, Wallace H Coulter Dept Biomed Engn, Atlanta, GA 30332 USA

[3] MIT, McGovern Inst Brain Res, Cambridge, MA 02139 USA

[4] MIT, Dept Brain & Cognit Sci, Cambridge, MA 02139 USA

[5] MIT, Dept Biol Engn, Cambridge, MA 02139 USA

[6] Harvard Univ, Sch Med, Dept Syst Biol, Boston, MA USA

来源：

NATURE BIOTECHNOLOGY | 2015年 / 33卷 / 11期

关键词：

GUIDE RNA; GENOME; CRISPR-CAS9; SPECIFICITY; COMPLEX; DNA; TRANSCRIPTION; ENDONUCLEASE; IMMUNITY; CELLS;

D O I：

10.1038/nbt.3390

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：

We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing doublestranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells.

引用

页码：1159 / U147

页数：4

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