RNA-programmed genome editing in human cells

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  • Martin Jinek
    Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States
  • Alexandra East
    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
  • Aaron Cheng
    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
  • Steven Lin
    Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States
  • Enbo Ma
    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States
  • Jennifer Doudna
    Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States

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Description

<jats:p>Type II CRISPR immune systems in bacteria use a dual RNA-guided DNA endonuclease, Cas9, to cleave foreign DNA at specific sites. We show here that Cas9 assembles with hybrid guide RNAs in human cells and can induce the formation of double-strand DNA breaks (DSBs) at a site complementary to the guide RNA sequence in genomic DNA. This cleavage activity requires both Cas9 and the complementary binding of the guide RNA. Experiments using extracts from transfected cells show that RNA expression and/or assembly into Cas9 is the limiting factor for Cas9-mediated DNA cleavage. In addition, we find that extension of the RNA sequence at the 3′ end enhances DNA targeting activity in vivo. These results show that RNA-programmed genome editing is a facile strategy for introducing site-specific genetic changes in human cells.</jats:p>

Journal

  • eLife

    eLife 2 e00471-, 2013-01-29

    eLife Sciences Publications, Ltd

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