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Orthogonal Cas9 proteins for RNA-guided gene regulation and editing

  • US 9,587,252 B2
  • Filed: 03/31/2015
  • Issued: 03/07/2017
  • Est. Priority Date: 07/10/2013
  • Status: Active Grant
First Claim
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1. A method of modulating expression of two or more target nucleic acids in a cell comprisingintroducing into the cell a first foreign nucleic acid encoding two or more guide RNAs complementary to the two or more target nucleic acids,introducing into the cell a second foreign nucleic acid encoding two or more orthogonal RNA guided nuclease null DNA binding proteins that respectively bind to the two or more target nucleic acids and are guided by the two or more guide RNAs,introducing into the cell a third foreign nucleic acid encoding two or more transcriptional regulator proteins or domains,wherein the two or more guide RNAs, the two or more orthogonal RNA guided nuclease-null DNA binding proteins, and the two or more transcriptional regulator proteins or domains are expressed,wherein two or more co-localization complexes form with each including a guide RNA, an orthogonal RNA guided nuclease null DNA binding protein, a transcriptional regulator protein or domain and a target nucleic acid and wherein the transcriptional regulator protein or domain regulates expression of the target nucleic acid.

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