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Methods Of Modifying A Sequence Using CRISPR

  • US 20160053272A1
  • Filed: 07/16/2015
  • Published: 02/25/2016
  • Est. Priority Date: 07/18/2014
  • Status: Abandoned Application
First Claim
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1. A method of modifying one or more target nucleic acid sequences comprising:

  • (a) contacting the one or more target nucleic acid sequences with;

    i) one or more ribonucleic acid (RNA) sequences wherein each RNA sequence comprises a portion that is complementary to all or a portion of one or more of the target nucleic acid sequences;

    ii) a CRISPR associated (Cas) protein having nuclease activity;

    iii) one or more exogenous nucleic acid sequences wherein at least one exogenous nucleic acid sequence comprises a 5′

    adapter sequence that hybridizes to a 5′

    flanking sequence of the target nucleic acid sequence and at least one exogenous nucleic acid sequence comprises a 3′

    adapter sequence that hybridizes to a 3′

    flanking sequence of the target nucleic acid sequence; and

    iv) a nucleic acid sequence that interacts with Cas protein;

    thereby producing a combination; and

    (b) maintaining the combination under conditions in which the one or more RNA sequences hybridize to all or the portion of the one or more target nucleic acid sequences to which each RNA sequence forms a complement thereby forming one or more base paired structures, and the one or more base paired structures and the nucleic acid sequence that interacts with Cas protein direct Cas protein to cleave the one or more target nucleic acid sequences;

    thereby modifying the one or more target nucleic acid sequences.

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