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CasX enzymes comprise a distinct family of RNA-guided genome editors.
Liu, Jun-Jie; Orlova, Natalia; Oakes, Benjamin L; Ma, Enbo; Spinner, Hannah B; Baney, Katherine L M; Chuck, Jonathan; Tan, Dan; Knott, Gavin J; Harrington, Lucas B; Al-Shayeb, Basem; Wagner, Alexander; Brötzmann, Julian; Staahl, Brett T; Taylor, Kian L; Desmarais, John; Nogales, Eva; Doudna, Jennifer A.
Affiliation
  • Liu JJ; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
  • Orlova N; California Institute for Quantitative Biosciences, University of California, Berkeley, CA, USA.
  • Oakes BL; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
  • Ma E; California Institute for Quantitative Biosciences, University of California, Berkeley, CA, USA.
  • Spinner HB; Innovative Genomics Institute, University of California, Berkeley, CA, USA.
  • Baney KLM; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
  • Chuck J; Innovative Genomics Institute, University of California, Berkeley, CA, USA.
  • Tan D; Innovative Genomics Institute, University of California, Berkeley, CA, USA.
  • Knott GJ; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
  • Harrington LB; Clayton Foundation Laboratories of Peptide Biology, Salk Institute for Biological Studies, La Jolla, CA, USA.
  • Al-Shayeb B; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
  • Wagner A; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
  • Brötzmann J; Department of Plant and Microbiology, University of California, Berkeley, CA, USA.
  • Staahl BT; Max-Planck-Institute for Biochemistry, Planegg, Germany.
  • Taylor KL; Faculty of Chemistry and Pharmacy, Ludwig-Maximilians-University, Munich, Germany.
  • Desmarais J; Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
  • Nogales E; Innovative Genomics Institute, University of California, Berkeley, CA, USA.
  • Doudna JA; Innovative Genomics Institute, University of California, Berkeley, CA, USA.
Nature ; 566(7743): 218-223, 2019 02.
Article in En | MEDLINE | ID: mdl-30718774
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.
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Full text: 1 Database: MEDLINE Main subject: CRISPR-Associated Proteins / CRISPR-Cas Systems / Gene Editing Limits: Humans Language: En Year: 2019 Type: Article
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PubMed Links
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Full text: 1 Database: MEDLINE Main subject: CRISPR-Associated Proteins / CRISPR-Cas Systems / Gene Editing Limits: Humans Language: En Year: 2019 Type: Article