Cas9
核糖核酸
生物
DNA
引导RNA
基因组编辑
RNA编辑
清脆的
计算生物学
基因组
非编码RNA
分子生物学
核酸内切酶
RNA沉默
细胞生物学
染色质
基因
遗传学
人类基因组
RNA干扰
作者
Martin Jinek,Alexandra East,Aaron M. Cheng,Steven H. Lin,Enbo Ma,Jennifer A. Doudna
出处
期刊:eLife
[eLife Sciences Publications, Ltd.]
日期:2013-01-29
卷期号:2
被引量:1676
摘要
Type II CRISPR immune systems in bacteria use a dual RNA-guided DNA endonuclease, Cas9, to cleave foreign DNA at specific sites. We show here that Cas9 assembles with hybrid guide RNAs in human cells and can induce the formation of double-strand DNA breaks (DSBs) at a site complementary to the guide RNA sequence in genomic DNA. This cleavage activity requires both Cas9 and the complementary binding of the guide RNA. Experiments using extracts from transfected cells show that RNA expression and/or assembly into Cas9 is the limiting factor for Cas9-mediated DNA cleavage. In addition, we find that extension of the RNA sequence at the 3′ end enhances DNA targeting activity in vivo. These results show that RNA-programmed genome editing is a facile strategy for introducing site-specific genetic changes in human cells.
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