Multiplex Genome Engineering Using CRISPR/Cas Systems

被引：11506

作者：

Cong, Le ^{[1
,2
,3
]}

Ran, F. Ann ^{[1
,2
,5
]}

Cox, David ^{[1
,2
,4
]}

Lin, Shuailiang ^{[1
,2
,6
]}

Barretto, Robert ^{[7
]}

Habib, Naomi ^{[1
,2
]}

Hsu, Patrick D. ^{[1
,2
,5
]}

Wu, Xuebing ^{[8
,9
]}

Jiang, Wenyan ^{[10
]}

Marraffini, Luciano A. ^{[10
]}

Zhang, Feng ^{[1
,2
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge Ctr 7, Cambridge, MA 02142 USA

[2] MIT, Dept Biol Engn, Dept Brain & Cognit Sci, McGovern Inst Brain Res, Cambridge, MA 02139 USA

[3] Harvard Univ, Sch Med, Program Biol & Biomed Sci, Boston, MA 02115 USA

[4] Harvard Univ, Sch Med, Harvard MIT Hlth Sci & Technol, Boston, MA 02115 USA

[5] Harvard Univ, Dept Mol & Cellular Biol, Cambridge, MA 02138 USA

[6] Tsinghua Univ, Sch Life Sci, Beijing 100084, Peoples R China

[7] Columbia Univ, Coll Phys & Surg, Dept Biochem & Mol Biophys, New York, NY 10032 USA

[8] MIT, Computat & Syst Biol Grad Program, Cambridge, MA 02139 USA

[9] MIT, Koch Inst Integrat Canc Res, Cambridge, MA 02139 USA

[10] Rockefeller Univ, Bacteriol Lab, New York, NY 10065 USA

来源：

SCIENCE | 2013年 / 339卷 / 6121期

关键词：

STREPTOCOCCUS-THERMOPHILUS; IMMUNE-SYSTEM; TAL EFFECTORS; CAS SYSTEMS; SMALL RNAS; DNA; BACTERIA; ENDONUCLEASE; NUCLEASES; DEFENSE;

D O I：

10.1126/science.1231143

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：

Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.

引用

页码：819 / 823

页数：5

共 29 条

[1] CRISPR-Cas Systems in Bacteria and Archaea: Versatile Small RNAs for Adaptive Defense and Regulation [J].