6E9F

EsCas13d-crRNA-target RNA ternary complex

Summary for 6E9F

• Entry DOI: 10.2210/pdb6e9f/pdb
• EMDB information: 9013 9014 9015
• Descriptor: EsCas13d, crRNA (52-MER), RNA (27-MER), ... (4 entities in total)
• Functional Keywords: crispr-cas, rnase, complex, rna binding protein-rna complex, rna binding protein/rna
• Biological source: [Eubacterium] siraeum DSM 15702; More
• Total number of polymer chains: 3
• Total formula weight: 135835.99

Authors

Zhang, C.,Lyumkis, D. (deposition date: 2018-08-01, release date: 2018-10-03, Last modification date: 2024-03-13)

Primary citation

Zhang, C.,Konermann, S.,Brideau, N.J.,Lotfy, P.,Wu, X.,Novick, S.J.,Strutzenberg, T.,Griffin, P.R.,Hsu, P.D.,Lyumkis, D.
Structural Basis for the RNA-Guided Ribonuclease Activity of CRISPR-Cas13d.
Cell, 175:212-223.e17, 2018

PubMed Abstract: CRISPR-Cas endonucleases directed against foreign nucleic acids mediate prokaryotic adaptive immunity and have been tailored for broad genetic engineering applications. Type VI-D CRISPR systems contain the smallest known family of single effector Cas enzymes, and their signature Cas13d ribonuclease employs guide RNAs to cleave matching target RNAs. To understand the molecular basis for Cas13d function and explain its compact molecular architecture, we resolved cryoelectron microscopy structures of Cas13d-guide RNA binary complex and Cas13d-guide-target RNA ternary complex to 3.4 and 3.3 Å resolution, respectively. Furthermore, a 6.5 Å reconstruction of apo Cas13d combined with hydrogen-deuterium exchange revealed conformational dynamics that have implications for RNA scanning. These structures, together with biochemical and cellular characterization, provide insights into its RNA-guided, RNA-targeting mechanism and delineate a blueprint for the rational design of improved transcriptome engineering technologies.

PubMed: 30241607
DOI: 10.1016/j.cell.2018.09.001

Experimental method

ELECTRON MICROSCOPY (3.3 Å)