Engineered meganucleases with recognition sequences found in the human T cell receptor alpha constant region gene

Patent metadata

Field	Value
Publication number	US-10799535-B2
Application number	US-201615766290-A
Country	US
Kind code	B2
Filing date	Oct 5, 2016
Priority date	Oct 5, 2015
Publication date	Oct 13, 2020
Grant date	Oct 13, 2020

Abstract

Disclosed herein are recombinant meganucleases engineered to recognize and cleave a recognition sequence present in the human T cell receptor alpha constant region gene. The present disclosure further relates to the use of such recombinant meganucleases in methods for producing genetically-modified eukaryotic cells.

First claim

The invention claimed is:

1. An engineered meganuclease that binds and cleaves at a recognition sequence consisting of SEQ ID NO:3, 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 wherein said second subunit binds to a second recognition half-site of said recognition sequence, and wherein said engineered meganuclease comprises an amino acid sequence having at least 96% identity to SEQ ID NO: 8.

2. The engineered meganuclease of claim 1 , wherein said first subunit comprises an amino acid sequence having at least 96% sequence identity to residues 198-344 of SEQ ID NO:8, and wherein said second subunit comprises an amino acid sequence having at least 96% sequence identity to residues 7-153 of SEQ ID NO:8.

3. The engineered meganuclease of claim 1 , wherein said first subunit comprises residues 198-344 of SEQ ID NO:8.

4. The engineered meganuclease of claim 1 , wherein said second subunit comprises residues 7-153 of SEQ ID NO:8.

5. The engineered meganuclease of claim 1 , wherein said engineered meganuclease comprises the amino acid sequence of SEQ ID NO:8.

Assignees

• Prec Biosciences Inc

Inventors

• Nicholson Michael G
• Smith James Jefferson
• Jantz Derek
• Bartsevich Victor
• Macleod Daniel T
• Antony Jeyaraj

Classifications

• A61K40/50

  characterised by the use of allogeneic cells · CPC title

• A61K40/4211

  CD19 or B4 · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K2239/31

  characterized by the route of administration · CPC title