期刊
SCIENCE
卷 337, 期 6096, 页码 816-821出版社
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.1225829
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资金
- HHMI
- Austrian Science Fund [W1207-B09]
- Univ. of Vienna
- Swedish Research Council [K2010-57X-21436-01-3, 621-2011-5752-LiMS]
- Kempe Foundation
- Umea University
- Austrian Science Fund (FWF) [W1207] Funding Source: Austrian Science Fund (FWF)
- Direct For Biological Sciences [950971] Funding Source: National Science Foundation
- Div Of Molecular and Cellular Bioscience [950971] Funding Source: National Science Foundation
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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