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TitleStructural Basis for the RNA-Guided Ribonuclease Activity of CRISPR-Cas13d.
Journal, issue, pagesCell, Vol. 175, Issue 1, Page 212-223.e17, Year 2018
Publish dateSep 20, 2018
AuthorsCheng Zhang / Silvana Konermann / Nicholas J Brideau / Peter Lotfy / Xuebing Wu / Scott J Novick / Timothy Strutzenberg / Patrick R Griffin / Patrick D Hsu / Dmitry Lyumkis /
PubMed AbstractCRISPR-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 ...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.
External linksCell / PubMed:30241607 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 6.5 Å
Structure data

EMDB-9013, PDB-6e9e:
EsCas13d-crRNA binary complex
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-9014, PDB-6e9f:
EsCas13d-crRNA-target RNA ternary complex
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-9015:
Apo-EsCas13d
Method: EM (single particle) / Resolution: 6.5 Å

Chemicals

ChemComp-MG:
Unknown entry

Source
  • unidentified bacterium (bacteria)
  • bacterium (bacteria)
  • [eubacterium] siraeum dsm 15702 (bacteria)
KeywordsRNA BINDING PROTEIN/RNA / CRISPR-Cas / RNase / Complex / RNA BINDING PROTEIN-RNA complex

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