Cas9-recombinase fusion proteins and uses thereof
AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, John Paul Guilinger, David B. Thompson
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Abstract
Some aspects of this disclosure provide compositions, methods, and kits for improving the specificity of RNA-programmable endonucleases, such as Cas9. Also provided are variants of Cas9, e.g., Cas9 dimers and fusion proteins, engineered to have improved specificity for cleaving nucleic acid targets. Also provided are compositions, methods, and kits for site-specific recombination, using Cas9 fusion proteins (e.g., nuclease-inactivated Cas9 fused to a recombinase catalytic domain). Such Cas9 variants are useful in clinical and research settings involving site-specific modification of DNA, for example, genomic modifications.
Claims7
A dimer or tetramer of a fusion protein wherein the fusion protein comprises two domains:
- (i) a nuclease-inactivated Cas9 (dCas9);
- and (ii) a recombinase catalytic domain;
- wherein the nuclease-inactivated Cas9 domain is bound to a guide RNA (gRNA);
- and
- wherein the dimer or the tetramer is bound to a nucleic acid.
Dependencies are inferred from the claim text.