Small molecule-triggered Cas9 protein with improved genome-editing specificity

被引:0
作者
Davis, Kevin M. [1 ,2 ]
Pattanayak, Vikram [3 ]
Thompson, David B. [1 ,2 ]
Zuris, John A. [1 ,2 ]
Liu, David R. [1 ,2 ]
机构
[1] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA
[2] Harvard Univ, Howard Hughes Med Inst, Cambridge, MA 02138 USA
[3] Massachusetts Gen Hosp, Dept Pathol, Boston, MA 02114 USA
基金
美国国家卫生研究院; 加拿大自然科学与工程研究理事会;
关键词
HUMAN-CELLS; NUCLEASE SPECIFICITY; DIRECTED EVOLUTION; MAMMALIAN-CELLS; RNA; NICKASES; CLEAVAGE; DELIVERY; INTEIN;
D O I
10.1038/NCHEMBIO.1793
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Directly modulating the activity of genome-editing proteins has the potential to increase their specificity by reducing activity following target locus modification. We developed Cas9 nucleases that are activated by the presence of a cell-permeable small molecule by inserting an evolved 4-hydroxytamoxifen-responsive intein at specific positions in Cas9. In human cells, conditionally active Cas9s modify target genomic sites with up to 25-fold higher specificity than wild-type Cas9.
引用
收藏
页码:316 / U97
页数:4
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