Fusion proteins comprising cas enzymes and dna-binding proteins
AssigneeSarepta Therapeutics, Inc.
InventorsEric WALTON, Ami M. KABADI
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Abstract
Disclosed herein are fusion protein comprising a Cas enzyme (e.g., a type II or a type IV Cas enzyme) or its fragment and a DNA-binding protein (e.g., a 7 kDa DNA-binding protein) or a DNA-binding protein fragment thereof, as well as the use of this fusion protein in order to enhance CRISPR-Cas systems genome-editing efficiency.
Claims136
A fusion protein comprising (a) a Cas enzyme or the fragment thereof and (b) a DNA-binding protein or a DNA-binding fragment thereof.
The fusion protein of any one of the preceding claims, wherein the DNA-binding protein or DNA-binding fragment thereof is located upstream of the Cas enzyme or its fragment, downstream of the Cas enzyme or its fragment, or a combination thereof.
The fusion protein of any one of the preceding claims, wherein the fusion protein further comprises at least one nuclear localization signal (NLS), at least one cell penetrating peptide, or a combination thereof.
The fusion protein of any one of the preceding claims, wherein the fusion protein has an editing efficiency that is increased compared to a same Cas enzyme or its fragment, lacking the DNA-binding protein or DNA-binding fragment thereof.
The fusion protein of any one of the preceding claims, wherein the Cas enzyme is a type II Cas enzyme or a type V Cas enzyme.
An expression cassette comprising a polynucleotide comprising a nucleotide sequence encoding a fusion protein, wherein the fusion protein comprises (1) a Cas enzyme or its fragment and (2) a DNA-binding protein or a DNA-binding fragment thereof.
The expression cassette of any one of the preceding claims, wherein the DNA-binding protein or DNA-binding fragment thereof is linked to the Cas enzyme or its fragment via one or more chemical bonds, by a linker, or a combination thereof.
The expression cassette of any one of the preceding claims, wherein the polynucleotide further comprises a nucleotide sequence encoding at least one nuclear localization signal (NLS), at least one cell penetrating peptide (CPP), or a combination thereof.
The expression cassette of any one of the preceding claims, wherein the polynucleotide further comprises a nucleotide sequence encoding a molecular tag, a selectable marker, a self-cleaving peptide or any combination thereof.
The expression cassette of any one of the preceding claims, wherein the polynucleotide further comprises a nucleotide sequence encoding a promoter, an enhancer, a silencer, a polyA tail, a poly adenylation signal, a 3′-UTR, a 5′-UTR, an intronic sequence, or any combination thereof.
The method of any one of the preceding claims, wherein the cell is in a subject.
Dependencies are inferred from the claim text.