5CZZ

Crystal structure of Staphylococcus aureus Cas9 in complex with sgRNA and target DNA (TTGAAT PAM)

5CZZ の概要

• エントリーDOI: 10.2210/pdb5czz/pdb
• 関連するPDBエントリー: 5AXW
• 分子名称: CRISPR-associated endonuclease Cas9, RNA (73-MER), DNA (28-MER), ... (8 entities in total)
• 機能のキーワード: crispr-cas9, genome engineering, hydrolase-rna-dna complex, hydrolase/rna/dna
• 由来する生物種: Staphylococcus aureus subsp. aureus; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 159161.08

構造登録者

Nishimasu, H.,Ishitani, R.,Nureki, O. (登録日: 2015-08-01, 公開日: 2015-09-02, 最終更新日: 2024-03-20)

主引用文献

Nishimasu, H.,Cong, L.,Yan, W.X.,Ran, F.A.,Zetsche, B.,Li, Y.,Kurabayashi, A.,Ishitani, R.,Zhang, F.,Nureki, O.
Crystal Structure of Staphylococcus aureus Cas9.
Cell, 162:1113-1126, 2015

PubMed Abstract: The RNA-guided DNA endonuclease Cas9 cleaves double-stranded DNA targets with a protospacer adjacent motif (PAM) and complementarity to the guide RNA. Recently, we harnessed Staphylococcus aureus Cas9 (SaCas9), which is significantly smaller than Streptococcus pyogenes Cas9 (SpCas9), to facilitate efficient in vivo genome editing. Here, we report the crystal structures of SaCas9 in complex with a single guide RNA (sgRNA) and its double-stranded DNA targets, containing the 5'-TTGAAT-3' PAM and the 5'-TTGGGT-3' PAM, at 2.6 and 2.7 Å resolutions, respectively. The structures revealed the mechanism of the relaxed recognition of the 5'-NNGRRT-3' PAM by SaCas9. A structural comparison of SaCas9 with SpCas9 highlighted both structural conservation and divergence, explaining their distinct PAM specificities and orthologous sgRNA recognition. Finally, we applied the structural information about this minimal Cas9 to rationally design compact transcriptional activators and inducible nucleases, to further expand the CRISPR-Cas9 genome editing toolbox.

PubMed: 26317473
DOI: 10.1016/j.cell.2015.08.007

実験手法

X-RAY DIFFRACTION (2.6 Å)