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US 2026/0250650 A1Published application
Base editing proteins and uses thereof
OwnerJohn KULMAN, Daniel FERULLO +3inventors
InventorsJohn KULMAN, Daniel FERULLO, Brian FOCHTMAN, Aishwarya VISHWANATHAN, Vasudevan ACHUTHAN
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Abstract
The present disclosure provides base editing fusion proteins, protein complexes, compositions, cells and methods of using the same for targeted nucleic acid editing. The base editing fusion protein can comprise a Cas9 domain and a deaminase domain capable of deaminating a nucleotide base in a target nucleic acid sequence.
Claims62
A fusion protein comprising:
- a Cas9 domain, wherein the Cas9 domain when associated with a guide RNA (gRNA) specifically binds to a target nucleic acid sequence;
- and
- a cytidine deaminase domain capable of deaminating a cytosine base in a single-stranded portion of the target nucleic acid sequence, wherein the cytidine deaminase domain comprises a deaminase from an apolipoprotein B mRNA-editing complex (APOBEC) family deaminase.
A method, comprising:
- transfecting a cell expressing an first Cas9 domain or a nucleic acid encoding the first Cas9 domain with a first guide RNA and a fusion protein comprising:
- (1) a second Cas9 domain, wherein the second Cas9 domain when associated with a second guide RNA specifically binds to a target nucleic acid sequence and (2) a cytidine deaminase domain capable of deaminating a cytosine base in a single-stranded portion of the target nucleic acid sequence, or a nucleic acid sequence encoding the fusion protein, wherein the first guide RNA when associated with the first Cas9 domain binds to a non-target nucleic acid sequence;
- and
- analyzing the genomic sequence of the cell.
1 more element
Dependencies are inferred from the claim text.