6NY2

CasX-gRNA-DNA(45bp) state I

Replaces:  6E79

Summary for 6NY2

• Entry DOI: 10.2210/pdb6ny2/pdb
• EMDB information: 8980 8994 8996
• Descriptor: DNA target strand, DNA Non-target strand, CasX, ... (4 entities in total)
• Functional Keywords: casx, sgrna, target dna, crispr, rna binding protein-rna-dna complex, rna binding protein/rna/dna
• Biological source: Deltaproteobacteria bacterium; More
• Total number of polymer chains: 4
• Total formula weight: 174931.92

Authors

Liu, J.J.,Orlova, N.,Nogales, E.,Doudna, J.A. (deposition date: 2019-02-10, release date: 2019-02-27, Last modification date: 2024-11-06)

Primary citation

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.
CasX enzymes comprise a distinct family of RNA-guided genome editors.
Nature, 566:218-223, 2019

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.

PubMed: 30718774
DOI: 10.1038/s41586-019-0908-x

Experimental method

ELECTRON MICROSCOPY (3.2 Å)