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Methods and compositions for RNA-directed target DNA modification and for RNA-directed modulation of transcription

  • US 10,487,341 B2
  • Filed: 02/15/2019
  • Issued: 11/26/2019
  • Est. Priority Date: 05/25/2012
  • Status: Active Grant
First Claim
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1. A composition comprising(a) a Cas9 protein or a nucleic acid encoding the Cas9 protein, wherein the Cas9 protein comprises a mutation in a RuvC and/or an HNH domain;

  • and(b) a single molecule DNA-targeting RNA or a nucleic acid encoding the single molecule DNA-targeting RNA, wherein the single molecule DNA-targeting RNA comprises in 5′

    to 3′

    order;

    (i) a targeter-RNA that is capable of hybridizing with a target sequence of a target DNA, and(ii) an activator-RNA that is capable of hybridizing with the targeter-RNA to form a double-stranded duplex,wherein (i) and (ii) are covalently linked by intervening nucleotides,wherein the single molecule DNA-targeting RNA is capable of forming a complex with the Cas9 protein and hybridization of the targeter-RNA to the target sequence is capable of targeting the Cas9 protein to the target DNA.

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