7N3P

Cryo-EM structure of the Cas12k-sgRNA-dsDNA complex

Summary for 7N3P

• Entry DOI: 10.2210/pdb7n3p/pdb
• EMDB information: 24143
• Descriptor: Cas12k, Single guide RNA, DNA (5'-D(*CP*AP*TP*GP*AP*CP*TP*TP*CP*TP*CP*AP*AP*CP*CP*GP*AP*GP*TP*TP*T)-3'), ... (4 entities in total)
• Functional Keywords: crispr cas, rna binding protein, protein-rna-dna complex, rna binding protein-rna-dna complex, rna binding protein/rna/dna
• Biological source: Scytonema hofmannii; More
• Total number of polymer chains: 4
• Total formula weight: 174153.07

Authors

Chang, L.,Li, Z.,Xiao, R.,Wang, S.,Han, R. (deposition date: 2021-06-01, release date: 2021-09-01, Last modification date: 2025-06-04)

Primary citation

Xiao, R.,Wang, S.,Han, R.,Li, Z.,Gabel, C.,Mukherjee, I.A.,Chang, L.
Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition.
Mol.Cell, 81:4457-4466.e5, 2021

PubMed Abstract: The type V-K CRISPR-Cas system, featured by Cas12k effector with a naturally inactivated RuvC domain and associated with Tn7-like transposon for RNA-guided DNA transposition, is a promising tool for precise DNA insertion. To reveal the mechanism underlying target DNA recognition, we determined a cryoelectron microscopy (cryo-EM) structure of Cas12k from cyanobacteria Scytonema hofmanni in complex with a single guide RNA (sgRNA) and a double-stranded target DNA. Coupled with mutagenesis and in vitro DNA transposition assay, our results revealed mechanisms for the recognition of the GGTT protospacer adjacent motif (PAM) sequence and the structural elements of Cas12k critical for RNA-guided DNA transposition. These structural and mechanistic insights should aid in the development of type V-K CRISPR-transposon systems as tools for genome editing.

PubMed: 34450043
DOI: 10.1016/j.molcel.2021.07.043

Experimental method

ELECTRON MICROSCOPY (3.65 Å)