IP Library › Granted Patent US 11,814,624
Granted Patent B2
US 11,814,624 · App. 17/498,531 · Granted Nov 14, 2023

Targeted non-viral DNA insertions

Inventors: Theodore Lee Roth (San Francisco, CA); Alexander Marson (San Francisco, CA)
Assignee: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
C12N15/1138C12N9/22C12N15/102C12N15/90C12N2310/20C12N2320/53
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Quick Facts
Patent No.
US 11,814,624
App. No.
17/498,531
Granted
Nov 14, 2023
Kind
B2
Abstract

Provided herein are methods and compositions for editing the genome of a cell. In some embodiments, a nucleotide sequence of at least 200 nucleotides in length is inserted into a target region in the genome of a cell.

Claims (34)

1. An isolated population of primary human immune cells, said population comprising one or more primary human immune cells that each comprise at least one heterologous DNA template non-virally inserted into a target region of the genome, wherein the size of the at least one heterologous DNA template is greater than or equal to 300 base pairs (bp).

2. The population of claim 1 , wherein the size of the at least one heterologous DNA template is greater than or equal to 1 kilobase (kb).

3. The population of claim 1 , wherein the size of the at least one heterologous DNA template is greater than or equal to 1.5 kb.

4. The population of claim 1 , wherein the size of the at least one heterologous DNA template is 300 bp to 5 kb.

5. The population of claim 1 , wherein the at least one heterologous DNA template is a double-stranded DNA template.

6. The population of claim 1 , wherein the at least one heterologous DNA template is a linear DNA template.

7. The population of claim 1 , wherein the primary human immune cells are primary human T cells.

8. The population of claim 1 , wherein the at least one heterologous DNA template comprises a gene.

9. The population of claim 1 , wherein the at least one heterologous DNA template encodes a chimeric antigen receptor (CAR).

10. The population of claim 1 , wherein the one or more primary human immune cells do not comprise a viral vector.

11. An isolated population of primary human immune cells, wherein at least 10% of the primary human immune cells in the population comprise at least one heterologous DNA template in a target region of the genome, wherein the size of the DNA template is greater than or equal to 300 base pairs, and wherein the primary human immune cells do not comprise a viral vector for introducing the at least one heterologous DNA template into the primary human immune cells.

12. The population of claim 11 , wherein the size of the at least one heterologous DNA template is greater than or equal to 1 kilobase (kb).

13. The population of claim 11 , wherein at least 20% of the primary human immune cells in the population comprise the at least one heterologous DNA template in the target region of the genome.

14. The population of claim 11 , wherein the size of the at least one heterologous DNA template is 300 bp to 5 kb.

15. The population of claim 11 , wherein the size of the at least one heterologous DNA template is greater than or equal to 500 bp.

16. The population of claim 11 , wherein the size of the at least one heterologous DNA template is 500 bp to 5 kb.

17. The population of claim 11 , wherein the size of the at least one heterologous DNA template is 1 kb to 5 kb.

18. The population of claim 11 , wherein the size of the at least one heterologous DNA template is greater than or equal to 1.5 kb.

19. The population of claim 11 , wherein the size of the at least one heterologous DNA template is 1.5 kb to 5 kb.

20. The population of claim 11 , wherein the primary human immune cells are primary human T cells.

21. The population of claim 11 , wherein the primary human immune cells comprise primary human cytotoxic T cells.

22. The population of claim 11 , wherein the primary human immune cells are primary human regulatory T cells.

23. The population of claim 11 , wherein the primary human immune cells are primary human NK cells.

24. A method of treating a disease in a human subject, the method comprising administering the population of primary human immune cells of claim 11 to the human subject.

25. A method of treating a disease in a human subject, the method comprising

administering the population of primary human immune cells of claim 1 to the human subject.

26. The isolated population of primary human immune cells of claim 1 , wherein at least 10% of the primary human immune cells in the population comprise the at least one heterologous DNA template non-virally inserted into the target region of the genome.

27. The population of claim 1 , wherein the size of the at least one heterologous DNA template is 1 kb to 5 kb.

28. The population of claim 1 , wherein the size of the at least one heterologous DNA template is 1.5 kb to 5 kb.

29. The population of claim 1 , wherein the size of the at least one heterologous DNA template is greater than or equal to 500 bp.

30. The population of claim 1 , wherein the size of the at least one heterologous DNA template is 500 bp to 5 kb.

31. The population of claim 1 , wherein the primary human immune cells comprise primary human cytotoxic T cells.

32. The population of claim 1 , wherein the primary human immune cells are primary human regulatory T cells.

33. The population of claim 1 , wherein the primary human immune cells are primary human NK cells.

Assignments (2)
CONFIRMATORY LICENSE Recorded Jan 10, 2024
From: UNIVERSITY OF CALIFORNIA SAN FRANCISCO
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 066254/0484 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 10, 2022
From: ROTH, THEODORE LEE; MARSON, ALEXANDER
To: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Reel/Frame 059225/0777 →
Continuity (4)
Continuation 16622843
Provisional Application 62552180 · Aug 30, 2017
Provisional Application 62520117 · Jun 15, 2017
Related Publication 20220064653A1 · Mar 3, 2022
Cited By (2)
US 12,234,461 US 12,257,304