6NY2

CasX-gRNA-DNA(45bp) state I


Experimental Data Snapshot

  • Method: ELECTRON MICROSCOPY
  • Resolution: 3.20 Å
  • Aggregation State: PARTICLE 
  • Reconstruction Method: SINGLE PARTICLE 

wwPDB Validation   3D Report Full Report


This is version 1.2 of the entry. See complete history


Literature

CasX enzymes comprise a distinct family of RNA-guided genome editors.

Liu, J.J.Orlova, N.Oakes, B.L.Ma, E.Spinner, H.B.Baney, K.L.M.Chuck, J.Tan, D.Knott, G.J.Harrington, L.B.Al-Shayeb, B.Wagner, A.Brotzmann, J.Staahl, B.T.Taylor, K.L.Desmarais, J.Nogales, E.Doudna, J.A.

(2019) Nature 566: 218-223

  • DOI: https://doi.org/10.1038/s41586-019-0908-x
  • Primary Citation of Related Structures:  
    6NY1, 6NY2, 6NY3

  • PubMed Abstract: 

    The RNA-guided CRISPR-associated (Cas) proteins Cas9 and Cas12a provide adaptive immunity against invading nucleic acids, and function as powerful tools for genome editing in a wide range of organisms. Here we reveal the underlying mechanisms of a third, fundamentally distinct RNA-guided genome-editing platform named CRISPR-CasX, which uses unique structures for programmable double-stranded DNA binding and cleavage. Biochemical and in vivo data demonstrate that CasX is active for Escherichia coli and human genome modification. Eight cryo-electron microscopy structures of CasX in different states of assembly with its guide RNA and double-stranded DNA substrates reveal an extensive RNA scaffold and a domain required for DNA unwinding. These data demonstrate how CasX activity arose through convergent evolution to establish an enzyme family that is functionally separate from both Cas9 and Cas12a.


  • Organizational Affiliation

    Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.


Macromolecules

Find similar proteins by:  (by identity cutoff)  |  3D Structure
Entity ID: 3
MoleculeChains Sequence LengthOrganismDetailsImage
CasXC [auth Y]986Deltaproteobacteria bacteriumMutation(s): 0 
Gene Names: DD725_10130
UniProt
Find proteins for A0A357BT59 (Deltaproteobacteria bacterium)
Explore A0A357BT59 
Go to UniProtKB:  A0A357BT59
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupA0A357BT59
Sequence Annotations
Expand
  • Reference Sequence
Find similar nucleic acids by:  (by identity cutoff)  |  3D Structure
Entity ID: 1
MoleculeChains LengthOrganismImage
DNA target strandA [auth C]45Deltaproteobacteria bacterium
Sequence Annotations
Expand
  • Reference Sequence
Find similar nucleic acids by:  (by identity cutoff)  |  3D Structure
Entity ID: 2
MoleculeChains LengthOrganismImage
DNA Non-target strandB [auth D]45Deltaproteobacteria bacterium
Sequence Annotations
Expand
  • Reference Sequence
Find similar nucleic acids by:  (by identity cutoff)  |  3D Structure
Entity ID: 4
MoleculeChains LengthOrganismImage
RNA (110-MER)D [auth B]122Deltaproteobacteria bacterium
Sequence Annotations
Expand
  • Reference Sequence
Experimental Data & Validation

Experimental Data

  • Method: ELECTRON MICROSCOPY
  • Resolution: 3.20 Å
  • Aggregation State: PARTICLE 
  • Reconstruction Method: SINGLE PARTICLE 

Structure Validation

View Full Validation Report



Entry History & Funding Information

Deposition Data


Funding OrganizationLocationGrant Number
National Institutes of Health/National Human Genome Research Institute (NIH/NHGRI)United Statesp01

Revision History  (Full details and data files)

  • Version 1.0: 2019-02-27
    Type: Initial release
  • Version 1.1: 2019-12-18
    Changes: Author supporting evidence
  • Version 1.2: 2019-12-25
    Changes: Database references