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Methods Of Depleting Target Sequences Using CRISPR

  • US 20160053304A1
  • Filed: 07/17/2015
  • Published: 02/25/2016
  • Est. Priority Date: 07/18/2014
  • Status: Abandoned Application
First Claim
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1. A method of depleting one or more target nucleic acid sequences in a sample comprising the one or more target nucleic acid sequences and one or more non-target nucleic acid sequences, wherein each of the target nucleic acid sequences and the non-target nucleic acid sequences comprise a 5′

  • adapter and a 3′

    adapter comprising;

    (a) contacting the sample with;

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

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

    iii) a nucleic acid sequence that interacts with the Cas protein;

    thereby producing a combination; and

    (b) maintaining the combination under conditions in which the RNA sequences are allowed to hybridize to all or a portion of the target nucleic acid sequence 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 the Cas protein direct the Cas protein to deplete each of the target nucleic acid sequences;

    thereby depleting the target nucleic acid in the sample.

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