8KAI

Crystal structure of SpyCas9-crRNA-tracrRNA complex bound to 17nt target DNA

8KAI の概要

• エントリーDOI: 10.2210/pdb8kai/pdb
• 分子名称: RNA (34-MER), CRISPR-associated endonuclease Cas9/Csn1, DNA (25-MER), ... (6 entities in total)
• 機能のキーワード: nuclease, rna binding protein/rna/dna, rna binding protein-rna-dna complex
• 由来する生物種: Streptococcus pyogenes serotype M1; 詳細
• タンパク質・核酸の鎖数: 10
• 化学式量合計: 402861.79

構造登録者

Chen, Y.,Chen, J.,Liu, L. (登録日: 2023-08-03, 公開日: 2024-06-05, 最終更新日: 2024-06-12)

主引用文献

Chen, J.,Chen, Y.,Huang, L.,Lin, X.,Chen, H.,Xiang, W.,Liu, L.
Trans-nuclease activity of Cas9 activated by DNA or RNA target binding.
Nat.Biotechnol., 2024

PubMed Abstract: Type V and type VI CRISPR-Cas systems have been shown to cleave nonspecific single-stranded DNA (ssDNA) or single-stranded RNA (ssRNA) in trans, but this has not been observed in type II CRISPR-Cas systems using single guide RNA. We show here that the type II CRISPR-Cas9 systems directed by CRISPR RNA and trans-activating CRISPR RNA dual RNAs show RuvC domain-dependent trans-cleavage activity for both ssDNA and ssRNA substrates. Cas9 possesses sequence preferences for trans-cleavage substrates, preferring to cleave T- or C-rich ssDNA substrates. We find that the trans-cleavage activity of Cas9 can be activated by target ssDNA, double-stranded DNA and ssRNA. The crystal structure of Cas9 in complex with guide RNA and target RNA provides a structural basis for the binding of target RNA to activate Cas9. Based on the trans-cleavage activity of Cas9 and nucleic acid amplification technology, we develop the nucleic acid detection platforms DNA-activated Cas9 detection and RNA-activated Cas9 detection, which are capable of detecting DNA and RNA samples with high sensitivity and specificity.

PubMed: 38811761
DOI: 10.1038/s41587-024-02255-7

実験手法

X-RAY DIFFRACTION (3.49 Å)