Genetic modification of the hydroxyacid oxidase 1 gene for treatment of primary hyperoxaluria
AssigneePrecision BioSciences, Inc.
InventorsRoshni Davey, Derek Jantz, James Jefferson Smith, Gary Owens
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Abstract
Disclosed are engineered nucleases that bind and cleave a recognition sequence within a hydroxyacid oxidase 1 (HAO1) gene. The present invention also encompasses methods of using such engineered nucleases to make genetically-modified cells. Further, the invention encompasses pharmaceutical compositions comprising engineered nuclease proteins or nucleic acids encoding engineered nucleases of the invention, and the use of such compositions for treatment of primary hyperoxaluria type I.
Claims134
An engineered meganuclease that binds and cleaves a recognition sequence comprising SEQ ID NO:
- 5 within an HAO1 gene, wherein said engineered meganuclease comprises a first subunit and a second subunit, wherein said first subunit binds to a first recognition half-site of said recognition sequence and comprises a first hypervariable (HVR1) region, and wherein said second subunit binds to a second recognition half-site of said recognition sequence and comprises a second hypervariable (HVR2) region.
A genetically-modified eukaryotic cell comprising a modified HAO1 gene, wherein said modified HAO1 gene encodes a modified HAO1 polypeptide which comprises the amino acids encoded by exons 1-7 of the HAO1 gene but lacks a peroxisomal targeting signal.
A method for producing a genetically-modified eukaryotic cell comprising a modified HAO1 gene, said method comprising introducing into a eukaryotic cell:
- (a) a nucleic acid encoding an engineered nuclease having specificity for a recognition sequence within an HAO1 gene, wherein said engineered nuclease is expressed in said eukaryotic cell;
- or
- (b) said engineered nuclease having specificity for a recognition sequence within an HAO1 gene;
- wherein said engineered nuclease produces a cleavage site within said recognition sequence and generates a modified HAO1 gene which encodes a modified HAO1 polypeptide, wherein said modified HAO1 polypeptide comprises the amino acids encoded by exons 1-7 of the HAO1 gene but lacks a peroxisomal targeting signal.
A pharmaceutical composition comprising a pharmaceutically acceptable carrier and:
- (a) a nucleic acid encoding an engineered nuclease having specificity for a recognition sequence within an HAO1 gene, wherein said engineered nuclease is expressed in a eukaryotic cell in vivo;
- or
- (b) said engineered nuclease having specificity for a recognition sequence within an HAO1 gene;
- wherein said engineered nuclease produces a cleavage site within said recognition sequence and generates a modified HAO1 gene which encodes a modified HAO1 polypeptide, wherein said modified HAO1 polypeptide comprises the amino acids encoded by exons 1-7 of the HAO1 gene but lacks a peroxisomal targeting signal.
A method for treating primary hyperoxaluria type I (PH1) in a subject in need thereof, said method comprising delivering to a target cell in said subject a nucleic acid encoding an engineered nuclease having specificity for a recognition sequence within an HAO1 gene, wherein said engineered nuclease is expressed in said target cell, wherein said engineered nuclease produces a cleavage site within said recognition sequence and generates a modified HAO1 gene which encodes a modified HAO1 polypeptide, wherein said modified HAO1 polypeptide comprises the amino acids encoded by exons 1-7 of the HAO1 gene but lacks a peroxisomal targeting signal.
A recombinant HAO1 polypeptide comprising the amino acids encoded by exons 1-7 of said HAO1 gene but lacking a functional peroxisomal targeting signal.
Dependencies are inferred from the claim text.