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

  • US 10,519,467 B2
  • Filed: 11/03/2017
  • Issued: 12/31/2019
  • Est. Priority Date: 05/25/2012
  • Status: Active Grant
First Claim
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1. A method of producing a genetically modified cell comprising:

  • (1) introducing into a target cell comprising a target DNA;

    (a) a Cas9 protein or a nucleic acid encoding the Cas9 protein; and

    (b) a DNA-targeting RNA that comprises;

    (i) a targeter-RNA comprising a nucleotide sequence that is complementary to, and hybridizes with, a target sequence of the target DNA, and(ii) an activator-RNA that hybridizes with the targeter-RNA to form a double-stranded RNA duplex,wherein the targeter-RNA and the activator-RNA are covalently linked,wherein said introducing results in modification of the target DNA thereby producing the genetically modified cell; and

    then(2) culturing said genetically modified cell to produce a population of cells.

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