Photoactivatable CRISPR-Cas9 for optogenetic genome editing

被引：475

作者：

Nihongaki, Yuta ^{[1
]}

Kawano, Fuun ^{[1
]}

Nakajima, Takahiro ^{[1
]}

Sato, Moritoshi ^{[1
]}

机构：

[1] Univ Tokyo, Grad Sch Arts & Sci, Meguro Ku, Tokyo, Japan

来源：

NATURE BIOTECHNOLOGY | 2015年 / 33卷 / 07期

基金：

日本学术振兴会;

关键词：

NUCLEASE-INDUCED MUTATIONS; HUMAN-CELLS; TARGET DNA; GUIDE RNA; CAS9; PROTEIN; SPECIFICITY; SYSTEM; ENDONUCLEASE; ACTIVATION;

D O I：

10.1038/nbt.3245

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 describe an engineered photoactivatable Cas9 (paCas9) that enables optogenetic control of CRISPR-Cas9 genome editing in human cells. paCas9 consists of split Cas9 fragments and photoinducible dimerization domains named Magnets. In response to blue light irradiation, paCas9 expressed in human embryonic kidney 293T cells induces targeted genome sequence modifications through both nonhomologous end joining and homology-directed repair pathways. Genome editing activity can be switched off simply by extinguishing the light. We also demonstrate activation of paCas9 in spatial patterns determined by the sites of irradiation. Optogenetic control of targeted genome editing should facilitate improved understanding of complex gene networks and could prove useful in biomedical applications.

引用

页码：755 / 760

页数：6

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