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US 9,803,194 B2Granted
Compositions and methods of nucleic acid-targeting nucleic acids
AssigneeCaribou Biosciences, Inc.
InventorsAndrew Paul May, Rachel E. Haurwitz, Jennifer A. Doudna, James M. Berger, Matthew Merrill Carter, Paul Daniel Donohoue
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Abstract
This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof.
Claims22
A method for identifying off-target binding sites relative to a target DNA sequence, comprising:
- contacting a mixture of DNA fragments comprising the target DNA sequence with a complex comprising an enzymatically inactive site-directed polypeptide encoded by a Cas gene and a nucleic acid-targeting nucleic acid, wherein (i) the complex comprises an affinity tag, and (ii) the complex binds to the target DNA sequence;
- purifying the DNA fragments bound by the complex using the affinity tag;
- and
- identifying the off-target binding sites based on the DNA fragments bound by the complex that do not comprise the target DNA binding sequence.
1 more element
Dependencies are inferred from the claim text.