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Multiplex genome engineering using CRISPR/Cas systems.

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      Abstract

      Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.

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      Affiliations
      [1 ] Broad Institute of MIT and Harvard, 7 Cambridge Center, Cambridge, MA 02142, USA.
      Journal
      Science
      Science (New York, N.Y.)
      American Association for the Advancement of Science (AAAS)
      1095-9203
      0036-8075
      Feb 15 2013
      : 339
      : 6121
      23287718 science.1231143 10.1126/science.1231143 3795411 NIHMS444192

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