NOVEL METHOD

• US 20160186207A1
• Filed: 12/17/2013
• Published: 06/30/2016
• Est. Priority Date: 12/17/2012
• Status: Active Application

• Current Assignee: The Babraham Institute
• Original Assignee: The Babraham Institute
• Inventors: REIK, Wolf, PEAT, Julian, HORE, Timothy, KRUEGER, Christel
• Application Number:
  US14/652,742
• Publication Number:
  US 20160186207A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/16 Otologicals
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2501/40 Regulators of development
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Y 114/11 with 2-oxoglutarate as one donor, and incorporation of one atom each of oxygen into both donors (1.14.11)

Abstract

The invention relates to a method of enhancing the potency of a cell (for example, to a totipotent state), by introducing a TET family gene, derivative or fragment thereof into the cell. The invention also relates to methods and kits for preparing cells with enhanced potency, and uses of said cells.

Claims

1. A method of enhancing the potency of a cell, wherein said method comprises the step of introducing a TET family gene, derivative or fragment thereof into the cell.

2. The method according to claim 1, wherein the cell is enhanced to a totipotent state.

3. The method according to claim 1, wherein the cell is enhanced to a pluripotent state, such as a true pluripotent state.

4. The method according to any one of claims 1 to 3, wherein the TET family gene, derivative or fragment thereof, is TET2 or TET3.

5. The method according to any one of claims 1 to 4, wherein the TET family gene, derivative or fragment thereof, is TET3.

6. The method according to any one of claims 1 to 5, wherein the TET family gene, derivative or fragment thereof, is a TET3 isoform of SEQ ID NO:

11 or 13.

7. The method according to any one of claims 1, 2 or 4 to 6, wherein the cell is a pluripotent cell, such as an embryonic stem (ES) cell, in particular an E14 embryonic stem (ES) cell.

8. The method according to any one of claims 1 to 6, wherein the cell is a somatic cell.

9. The method according to any one of claims 1 to 8, wherein the introducing step comprises transfecting the cell with a vector containing the TET family gene, derivative or fragment thereof.

10. The method according to claim 9, wherein the vector is a transposon vector.

18. A cell with enhanced potency obtainable by the method defined in any one of claims 1 to 17.

22. The cell with enhanced potency according to claim 18 for use in therapy.

23. The cell with enhanced potency for use according to claim 22, wherein the therapy comprises tissue regeneration.

24. A kit comprising a vector containing a TET family gene, derivative or fragment thereof and instructions to use said kit in accordance with the method defined in any one of claims 1 to 17.

25. The kit according to claim 24, additionally comprising at least one pluripotent cell, such as an embryonic stem (ES) cell, in particular an E14 embryonic stem (ES) cell.

26. The kit according to claim 24, additionally comprising at least one somatic cell.

27. The kit according to claim 25 or claim 26, additionally comprising a medium for culturing the cell and instructions for preparing the cells with enhanced potency in accordance with the method defined in any one of claims 1 to 17.

11. A method of preparing a cell with enhanced potency which comprises the step of introducing a TET family gene, derivative or fragment thereof into a cell.

12. The method according to claim 11, therein the cell is a pluripotent cell, such as an embryonic stem (ES) cell, in particular an E14 embryonic stem (ES) cell.

13. The method according to claim 11, wherein the cell is a somatic cell.

14. The method according to claim 13, further comprising the step of introducing a Oct3/4 gene, a Sox2 gene, a Klf4 gene and a c-Myc gene into the somatic cell.

15. The method according to any one of claims 11 to 14, further comprising the step of culturing the cell after introduction of the TET family gene, derivative or fragment thereof.

16. The method according to any one of claims 11 to 15, further comprising the step of selecting one or more cells which overexpress the TET family gene, derivative or fragment thereof.

17. The method according to claim 16, wherein the one or more cells are selected using flow cytometry.

19. A nucleic acid comprising a TET3 isoform of SEQ ID NO:

11 or 13.

20. A vector comprising the nucleic acid according to claim 19.

21. Use of the nucleic acid according to claim 19, or the vector according to claim 20 in a method of enhancing the potency of a cell.