Composition and use of Cas protein inhibitors
AssigneeShanghaitech University
InventorsJia Liu, Biao Jiang, Peixiang Ma, Guang Yang
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Abstract
Provided are amino acid sequences capable of binding to and inhibiting a Cas protein's ability to bind to a nucleic acid molecule, thereby inhibiting the Cas protein's function in genome editing. Such Cas protein inhibitors, which can be comprised of a major coat protein (G8P), an extracellular region of the G8P (G8PEX), or a biological equivalent, are useful in improving the specificity of Cas protein-based genome editing procedures.
Claims13
A method for improving the specificity of a Cas protein-based genome editing procedure, comprising introducing to a cell undergoing the Cas protein-based genome editing procedure a non-bacteriophage vector comprising a polynucleotide encoding a polypeptide comprising a bacteriophage major coat protein G8P, an extracellular region of the G8P (G8PEX), or an amino acid sequence having at least 70% sequence identity to the G8P or the G8PEX and capable of binding to the Cas protein, wherein the Cas protein is a Cas9 protein or a Cas12a protein, and wherein the polynucleotide further comprises an inducible promoter regulating the expression of the polypeptide.
A method of genome editing in a subject, comprising administering to the subject a Cas protein-based genome editing system and a non-bacteriophage vector comprising a polynucleotide encoding a polypeptide operatively linked to an inducible promoter, wherein the polypeptide comprises a bacteriophage major coat protein G8P, an extracellular region of the G8P (G8PEX), or an amino acid sequence having at least 70% sequence identity to the G8P or the G8PEX and capable of binding to the Cas protein, wherein the Cas protein is a Cas9 protein or a Cas12a protein.
Dependencies are inferred from the claim text.