期刊
CELL
卷 156, 期 5, 页码 935-949出版社
CELL PRESS
DOI: 10.1016/j.cell.2014.02.001
关键词
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资金
- PRESTO from the Japan Science and Technology (JST) Agency and Platform for Drug Discovery and Informatics
- Japan Society for the Promotion of Science (JSPS) through its Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST Program)
- Core Research for Evolutional Science and Technology (CREST) Program, The Creation of Basic Medical Technologies to Clarify and Control the Mechanisms Underlying Chronic Inflammation of Japan Science and Technology Agency (JST)
- NIH Director's Pioneer Award [5DP1-MH100706]
- Keck Foundation
- McKnight Foundation
- Poitras Foundation
- Merkin Foundation
- Vallee Foundation
- Damon Runyon Foundation
- Searle Scholars Foundation
- Klingenstein Foundation
- Simons Foundation
- Grants-in-Aid for Scientific Research [23687014, 25116705] Funding Source: KAKEN
The CRISPR-associated endonuclease Cas9 can be targeted to specific genomic loci by single guide RNAs (sgRNAs). Here, we report the crystal structure of Streptococcus pyogenes Cas9 in complex with sgRNA and its target DNA at 2.5 angstrom resolution. The structure revealed a bilobed architecture composed of target recognition and nuclease lobes, accommodating the sgRNA: DNA heteroduplex in a positively charged groove at their interface. Whereas the recognition lobe is essential for binding sgRNA and DNA, the nuclease lobe contains the HNH and RuvC nuclease domains, which are properly positioned for cleavage of the complementary and noncomplementary strands of the target DNA, respectively. The nuclease lobe also contains a carboxyl-terminal domain responsible for the interaction with the protospacer adjacent motif (PAM). This high-resolution structure and accompanying functional analyses have revealed the molecular mechanism of RNA-guided DNA targeting by Cas9, thus paving the way for the rational design of new, versatile genome-editing technologies.
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