IP Library › Granted Patent US 12,264,343
Granted Patent B2
US 12,264,343 · App. 18/174,165 · Granted Apr 1, 2025

TCRa homing endonuclease variants

Inventors: Jordan Jarjour (Seattle, WA); Mark Pogson (North Bend, WA)
Assignee: REGENERON PHARMACEUTICALS, INC.
C12N9/22C12N15/85
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Quick Facts
Patent No.
US 12,264,343
App. No.
18/174,165
Granted
Apr 1, 2025
Kind
B2
Abstract

The present disclosure provides improved genome editing compositions and methods for editing a TCRα gene.

Claims (23)

1. A method of editing a genome of a cell, comprising: introducing into the cell one or more polynucleotides comprising a nucleotide sequence encoding a polypeptide comprising an I-OnuI homing endonuclease (HE) variant that cleaves a target site in the human T-cell receptor alpha (TCRα) gene, wherein the I-OnuI HE variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 7 or SEQ ID NO: 8.

2. The method of claim 1 , wherein the I-OnuI HE variant comprises the amino acid sequence set forth in SEQ ID NO: 7 or SEQ ID NO: 8.

3. The method of claim 1 , wherein the polypeptide binds and cleaves said TCRα gene at the polynucleotide sequence set forth in SEQ ID NO: 17.

4. The method of claim 1 , wherein the polypeptide further comprises a TALE DNA binding domain.

5. The method of claim 4 , wherein the TALE DNA binding domain comprises 11.5 TALE repeat units, and wherein the TALE DNA binding domain binds the polynucleotide sequence set forth in SEQ ID NO: 19.

6. The method of claim 4 , wherein the TALE DNA binding domain comprises 10.5 TALE repeat units, and wherein the TALE DNA binding domain binds the polynucleotide sequence set forth in SEQ ID NO: 18.

7. The method of claim 4 , wherein the polypeptide binds and cleaves the TCRα gene at the polynucleotide sequence set forth in SEQ ID NO: 20.

8. The method of claim 1 , wherein the polypeptide further comprises:

(a) a peptide linker or a viral self-cleaving 2A peptide; and

(b) a 3′-5′ exonuclease.

9. The method of claim 4 , wherein the polypeptide further comprises:

(a) a peptide linker or a viral self-cleaving 2A peptide; and

(b) a 3′-5′ exonuclease.

10. The method of claim 8 , wherein the 3′-5′ exonuclease is Trex2.

11. The method of claim 9 , wherein the 3′-5′ exonuclease is Trex2.

12. The method of claim 1 , wherein a vector comprises the polynucleotide.

13. The method of claim 1 , wherein the cell is a hematopoietic cell.

14. The method of claim 1 , wherein the cell is a T-cell or immune effector cell.

15. A method of editing a genome of a cell, comprising: introducing into the cell one or more polynucleotides comprising a nucleotide sequence encoding a polypeptide comprising an I-OnuI homing endonuclease (HE) variant that cleaves a target site in the human T-cell receptor alpha (TCRα) gene and further comprising a TALE DNA binding domain, wherein the polypeptide comprises an amino acid sequence set forth in SEQ ID NO: 10 or an amino acid sequence that is at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 12.

16. The method of claim 15 , wherein the polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 10 or SEQ ID NO: 12.

17. The method of claim 15 , wherein a vector comprises the one or more polynucleotides.

18. The method of claim 15 , wherein the cell is a hematopoietic cell.

19. The method of claim 15 , wherein the cell is a T-cell or immune effector cell.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 25, 2024
From: 2SEVENTY BIO, INC.
To: REGENERON PHARMACEUTICALS, INC.
Reel/Frame 067822/0847 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 8, 2023
From: BLUEBIRD BIO, INC.
To: 2SEVENTY BIO, INC.
Reel/Frame 064522/0806 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 8, 2023
From: JARJOUR, JORDAN; POGSON, MARK
To: BLUEBIRD BIO, INC.
Reel/Frame 064523/0302 →
Continuity (4)
Continuation 16340257
Provisional Application 62414266 · Oct 28, 2016
Provisional Application 62406689 · Oct 11, 2016
Related Publication 20230357736A1 · Nov 9, 2023
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