Cas variants for gene editing
AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, Alexis Christine Komor
Life of this patent
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Abstract
Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. In some embodiments, fusion proteins of Cas9 and nucleic acid editing enzymes or enzyme domains, e.g., deaminase domains, are provided. In some embodiments, methods for targeted nucleic acid editing are provided. In some embodiments, reagents and kits for the generation of targeted nucleic acid editing proteins, e.g., fusion proteins of Cas9 and nucleic acid editing enzymes or domains, are provided.
Claims37
A multi-molecular complex comprising:
- (a) a cytidine deaminase;
- (b) a Cas9 variant protein comprising the amino acid sequence of SEQ ID NO:
- 2 or SEQ ID NO:
- 37, or an amino acid sequence having at least 90% sequence identity with SEQ ID NO:
- 2 or SEQ ID NO:
- 37, wherein the Cas9 variant comprises one or more nuclease inactivating mutations within a RuvC1 subdomain and/or a HNH subdomain;
- and
- (c) a single guide RNA (sgRNA) comprising a region complementary to a strand of the target nucleic acid molecule;
- wherein the cytidine deaminase, the target nucleic acid molecule, the Cas9 variant, and the sgRNA associate in a multi-molecular complex;
- and
- wherein the multi-molecular complex deaminates a cytidine in the target nucleic acid molecule.
Dependencies are inferred from the claim text.