7YOJ

Structure of CasPi with guide RNA and target DNA

Summary for 7YOJ

• Entry DOI: 10.2210/pdb7yoj/pdb
• EMDB information: 33983
• Descriptor: CasPi, DNA (5'-D(P*CP*GP*GP*GP*AP*TP*GP*CP*CP*CP*AP*G)-3'), DNA (30-MER), ... (4 entities in total)
• Functional Keywords: caspi complex, rna binding protein, rna binding protein-dna-rna complex, rna binding protein/dna/rna
• Biological source: Armatimonadota bacterium; More
• Total number of polymer chains: 4
• Total formula weight: 168024.22

Authors

Li, C.P.,Wang, J.,Liu, J.J. (deposition date: 2022-08-01, release date: 2023-02-15, Last modification date: 2024-07-03)

Primary citation

Sun, A.,Li, C.P.,Chen, Z.,Zhang, S.,Li, D.Y.,Yang, Y.,Li, L.Q.,Zhao, Y.,Wang, K.,Li, Z.,Liu, J.,Liu, S.,Wang, J.,Liu, J.G.
The compact Cas pi (Cas12l) 'bracelet' provides a unique structural platform for DNA manipulation.
Cell Res., 33:229-244, 2023

PubMed Abstract: CRISPR-Cas modules serve as the adaptive nucleic acid immune systems for prokaryotes, and provide versatile tools for nucleic acid manipulation in various organisms. Here, we discovered a new miniature type V system, CRISPR-Casπ (Cas12l) (~860 aa), from the environmental metagenome. Complexed with a large guide RNA (~170 nt) comprising the tracrRNA and crRNA, Casπ (Cas12l) recognizes a unique 5' C-rich PAM for DNA cleavage under a broad range of biochemical conditions, and generates gene editing in mammalian cells. Cryo-EM study reveals a 'bracelet' architecture of Casπ effector encircling the DNA target at 3.4 Å resolution, substantially different from the canonical 'two-lobe' architectures of Cas12 and Cas9 nucleases. The large guide RNA serves as a 'two-arm' scaffold for effector assembly. Our study expands the knowledge of DNA targeting mechanisms by CRISPR effectors, and offers an efficient but compact platform for DNA manipulation.

PubMed: 36650285
DOI: 10.1038/s41422-022-00771-2

Experimental method

ELECTRON MICROSCOPY (3.36 Å)