SNAP output for PDB entry 4oo8 [SNAP web server]

PDB-id

4oo8; SNAP-derived features in text and JSON formats; DNAproDB

Class

hydrolase-DNA-RNA

Method

X-ray (2.5 Å)

Summary

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

Reference

Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O (2014): "Crystal structure of Cas9 in complex with guide RNA and target DNA." Cell(Cambridge,Mass.), 156, 935-949. doi: 10.1016/j.cell.2014.02.001.

Abstract

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 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.