反式激活crRNA
清脆的
Cas9
生物
DNA
核酸内切酶
基因组编辑
引导RNA
质粒
计算生物学
核糖核酸
遗传学
核酸酶
基因
作者
Martin Jinek,Krzysztof Chylinski,Ines Fonfara,M. Hauer,Jennifer A. Doudna,Emmanuelle Charpentier
出处
期刊:Science
[American Association for the Advancement of Science (AAAS)]
日期:2012-08-17
卷期号:337 (6096): 816-821
被引量:11884
标识
DOI:10.1126/science.1225829
摘要
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.
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