Treatment of retinitis pigmentosa using improved engineered meganucleases

Patent metadata

Field	Value
Publication number	US-12595469-B2
Application number	US-202117924352-A
Country	US
Kind code	B2
Filing date	May 11, 2021
Priority date	May 12, 2020
Publication date	Apr 7, 2026
Grant date	Apr 7, 2026

Abstract

Disclosed are recombinant meganucleases engineered to bind and cleave a recognition sequence present in a mutant RHO P23H allele. The invention further relates to the use of such recombinant meganucleases in a method for treating retinitis pigmentosa, wherein the mutant RHO P23H allele is preferentially targeted, cleaved, and inactivated.

First claim

What is claimed is:

1 . An engineered meganuclease that binds and cleaves a recognition sequence consisting of SEQ ID NO: 7 in a rhodopsin 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, wherein said second subunit binds to a second recognition half-site of said recognition sequence and comprises a second hypervariable (HVR2) region, and wherein said engineered meganuclease comprises the amino acid sequence of SEQ ID NO: 11.

2 . A polynucleotide comprising a nucleic acid sequence encoding said engineered meganuclease of claim 1 .

3 . A recombinant DNA construct comprising a polynucleotide comprising a nucleic acid sequence encoding said engineered meganuclease of claim 1 .

4 . The recombinant DNA construct of claim 3 , wherein said recombinant DNA construct encodes a recombinant virus comprising said polynucleotide.

5 . The recombinant DNA construct of claim 4 , wherein said recombinant virus is a recombinant adeno-associated virus (AAV).

6 . The recombinant DNA construct of claim 5 , wherein said recombinant AAV has an AAV5 or AAV2 serotype.

7 . The recombinant DNA construct of claim 3 , wherein said nucleic acid sequence comprises an eye-specific promoter sequence operably linked to said nucleic acid sequence encoding said engineered meganuclease.

8 . The recombinant DNA construct of claim 7 , wherein said promoter is a human G-protein-coupled receptor protein kinase 1 (GRK1) promoter.

9 . A recombinant virus comprising a polynucleotide comprising a nucleic acid sequence encoding said engineered meganuclease of claim 1 .

10 . The recombinant virus of claim 9 , wherein said recombinant virus is a recombinant adenovirus or a recombinant AAV.

11 . The recombinant virus of claim 10 , wherein said recombinant AAV has an AAV5 or AAV2 serotype.

12 . The recombinant virus of claim 9 , wherein said nucleic acid sequence comprises an eye-specific promoter sequence operably linked to said nucleic acid sequence encoding said engineered meganuclease.

13 . The recombinant virus of claim 12 , wherein said promoter is a human G-protein-coupled receptor protein kinase 1 (GRK1) promoter.

14 . A lipid nanoparticle composition comprising lipid nanoparticles comprising a polynucleotide, wherein said polynucleotide comprises a nucleic acid sequence encoding said engineered meganuclease of claim 1 .

Assignees

• Prec Biosciences Inc

Inventors

• Bartsevich Victor
• Jantz Derek
• Smith James Jefferson
• Nicholson Michael G

Classifications

• C12N2800/80

  Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites · CPC title

• C12N2750/14145

  Special targeting system for viral vectors · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N15/907

  in mammalian cells · CPC title

• C12N15/86

  Viral vectors · CPC title