4OO8

Crystal structure of Streptococcus pyogenes Cas9 in complex with guide RNA and target DNA

4OO8 の概要

• エントリーDOI: 10.2210/pdb4oo8/pdb
• 分子名称: CRISPR-associated endonuclease Cas9/Csn1, RNA (97-MER), DNA (5'-D(*CP*CP*AP*GP*CP*CP*AP*AP*GP*CP*GP*CP*AP*CP*CP*TP*AP*AP*TP*TP*TP*CP*C)-3'), ... (4 entities in total)
• 機能のキーワード: hydrolase-dna-rna complex, hydrolase/dna/rna
• 由来する生物種: Streptococcus pyogenes serotype M1
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 395168.71

構造登録者

Nishimasu, H.,Ishitani, R.,Nureki, O. (登録日: 2014-01-31, 公開日: 2014-02-26, 最終更新日: 2024-04-03)

主引用文献

Nishimasu, H.,Ran, F.A.,Hsu, P.D.,Konermann, S.,Shehata, S.I.,Dohmae, N.,Ishitani, R.,Zhang, F.,Nureki, O.
Crystal structure of Cas9 in complex with guide RNA and target DNA
Cell(Cambridge,Mass.), 156:935-949, 2014

PubMed 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.

PubMed: 24529477
DOI: 10.1016/j.cell.2014.02.001

実験手法

X-RAY DIFFRACTION (2.5 Å)