Granted Patent
B2
US 12,227,740 · App. 16/616,713 · Granted Feb 18, 2025
CBLB endonuclease variants, compositions, and methods of use
Inventors:
Jordan Jarjour (Seattle, WA); Kyle Havens (Seattle, WA); Anne-Rachel Krostag (Lake Forest Park, WA)
Assignee:
REGENERON PHARMACEUTICALS, INC.
C12N15/102A61K39/4611A61K39/4631A61K39/464404A61P35/00C07K16/2863C12N5/0634C12N5/0636C12N5/0646C12N9/10C12N9/22C12N15/907A61K2239/38C07K2319/80C07K2319/81C12N2510/00C12N2800/80
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Abstract
The present disclosure provides improved genome editing compositions and methods for editing a CBLB gene. The disclosure further provides genome edited cells for the prevention, treatment, or amelioration of at least one symptom of, a cancer, an infectious disease, an autoimmune disease, an inflammatory disease, or an immunodeficiency.
Claims (15)
1. A method of editing a human CBLB gene in a cell comprising:
introducing a polynucleotide encoding a polypeptide into the cell, wherein the polynucleotide encodes an I-OnuI homing endonuclease (HE) variant comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12;
wherein the I-OnuI HE variant binds and cleaves a target site in the human casitas B-lineage (Cbl) lymphoma proto-oncogene B (CBLB) gene set forth in SEQ ID NO: 20,
wherein the cleaving is at the nucleotides ATTC in SEQ ID NO: 20 and creates a double strand break, and
wherein the break is repaired by non-homologous end joining (NHEJ).
2. The method of claim 1 , wherein the cell is a hematopoietic cell.
3. The method of claim 1 , wherein the polynucleotide encoding the polypeptide is DNA.
4. The method of claim 1 , wherein the polynucleotide encoding the polypeptide is an mRNA.
5. The method of claim 1 , wherein the polynucleotide is introduced into the cell by electroporation.
6. The method of claim 1 , wherein the polynucleotide is introduced by lipid nanoparticle.
7. The method of claim 1 , wherein the I-OnuI HE variant comprises the amino acid sequence that is 100% identical to SEQ ID NO: 6.
8. The method of claim 1 , wherein the cell is a cytotoxic T lymphocyte (CTL), a tumor infiltrating lymphocyte (TIL), a helper T cell, a natural killer (NK) cell, or a natural killer T (NKT) cell.
9. The method of claim 1 , wherein the cell is a T cell.
10. The method of claim 1 , wherein the cell is a CD3 + , CD4 + , and/or CD8 + cell.
11. The method of claim 1 , wherein the cell is an immune effector cell.
Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 25, 2024
From: 2SEVENTY BIO, INC.
To: REGENERON PHARMACEUTICALS, INC.
Reel/Frame 067822/0416 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Sep 14, 2021
From: BLUEBIRD BIO, INC.
To: 2SEVENTY BIO, INC.
Reel/Frame 057683/0099 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Mar 4, 2020
From: JARJOUR, JORDAN; HAVENS, KYLE; KROSTAG, ANNE-RACHEL
To: BLUEBIRD BIO, INC.
Reel/Frame 052003/0234 →
Continuity (3)
Provisional Application
62511194
· May 25, 2017
Provisional Application
62567417
· Oct 3, 2017
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