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US 12,188,059 B2Granted
Proximity-dependent biotinylation and uses thereof
AssigneeThe Broad Institute, Inc.
InventorsSteven A. Carr, Jason Wright, Sam Myers
Life of this patent
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Abstract
The present invention provides fusion proteins, polynucleotides, kits, as well as TALE- or CRISPR-Cas based systems and methods. The present invention relies on proximity-dependent biotinylation, which allows site-directed protein or DNA purification and identification. The present invention provides tools for delineating the genetics of disease mechanism and for the identification of therapeutic targets and markers.
Claims26
A fusion protein comprising:
- (a) a nucleotide-editing protein, wherein the nucleotide-editing protein is a Streptococcus pyogenes or Staphylococcus aureus catalytically-inactive Cas9 (dCas9) protein, and
- (b) a proximity-dependent protein to catalyze ligation of biotin, or an ATP-biotin interaction loop or a biotin pocket thereof, wherein said proximity-dependent protein is a mutated E. coli Bir A, or a biotin-peroxide dependent ascorbate peroxidase (APEX) or a Split Biotin protein ligase separated at amino-acid positions (1-149, 141-320).
Dependencies are inferred from the claim text.