Scientific Publications

Crystal structure of cas9 in complex with guide RNA and target DNA.

Publication TypeJournal Article
AuthorsNishimasu, H., Ran FA, Hsu PD, Konermann S., Shehata SI, Dohmae N., Ishitani R., Zhang F., and Nureki O.
AbstractThe 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 Å 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.
Year of Publication2014
JournalCell
Volume156
Issue5
Pages935-49
Date Published (YYYY/MM/DD)2014/02/27
ISSN Number0092-8674
DOI10.1016/j.cell.2014.02.001
PubMedhttp://www.ncbi.nlm.nih.gov/pubmed/24529477?dopt=Abstract