METHODS FOR REPROGRAMMING SOMATIC CELLS

• US 20100144031A1
• Filed: 02/09/2010
• Published: 06/10/2010
• Est. Priority Date: 11/26/2003
• Status: Active Grant

• Current Assignee: Whitehead Institute for Biomedical Research
• Original Assignee: Whitehead Institute for Biomedical Research
• Inventors: Jaenisch, Rudolf, Hochedlinger, Konrad
• Granted Patent: US 8,940,536 B2
• Time in Patent Office: Days
• US Class Current: 435/354
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 67/0273 Cloned vertebrates
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 15/8775 Murine embryos
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/605 Nanog
• CPC Class Codes: C12N 2506/1307 from adult fibroblasts
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2830/003 tet inducible
• CPC Class Codes: C12N 2830/006 tet repressible
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The invention provides methods for reprogramming somatic cells to generate multipotent or pluripotent cells. Such methods are useful for a variety of purposes, including treating or preventing a medical condition in an individual. The invention further provides methods for identifying an agent that reprograms somatic cells to a less differentiated state.

Claims

1. A method of reprogramming a primary somatic cell to a less differentiated state, comprising:

introducing at least one exogenous pluripotency gene operably linked to at least one regulatory sequence into the somatic cell, wherein expression of the exogenously introduced nucleic acid results in expression of an endogenous pluripotency gene, thereby reprogramming the somatic cell to a less differentiated state.

2. The method of claim 1, wherein the exogenous pluripotency gene encodes Oct4, Nanog, or Sox2.

3. The method of claim 1, wherein the endogenous pluripotency gene encodes Oct4 or Nanog.

4. The method of claim 1, wherein the somatic cell does not comprise a selectable marker integrated into the endogenous locus of the pluripotency gene.

5. The method of claim 1, wherein the somatic cell is a human cell or a mouse cell.

6. The method of claim 1, wherein the somatic cell is an adult stem cell.

7. The method of claim 6, wherein the adult stem cell is a hematopoietic stem cell, neural stem cell, or mesenchymal stem cell.

8. A method of reprogramming a primary somatic cell to a less differentiated state, comprising:

(a) contacting the somatic cell with a candidate agent of interest with respect to its potential to reprogram a somatic cell; and

(b) introducing at least one exogenous pluripotency gene operably linked to at least one regulatory sequence into the somatic cell, wherein expression of the exogenously introduced pluripotency gene results in expression of an endogenous pluripotency gene, thereby reprogramming the somatic cell to a less differentiated state.

9. The method of claim 8, wherein the exogenous pluripotency gene encodes Oct4, Nanog, or Sox2.

10. The method of claim 8, wherein the endogenous pluripotency gene encodes Oct4 or Nanog.

11. The method of claim 8, wherein the somatic cell does not comprise a selectable marker integrated into the endogenous locus of the pluripotency gene.

12. The method of claim 8, wherein the somatic cell is a human cell or a mouse cell.

13. The method of claim 8, wherein the somatic cell is an adult stem cell.

14. The method of claim 13, wherein the adult stem cell is a hematopoietic stem cell, neural stem cell, or mesenchymal stem cell.

15. The method of claim 8, wherein the agent is a DNA methylation inhibitor, a histone deacetylase inhibitor or PD098059.

16. The method of claim 8, wherein the agent is a pluripotency protein selected from the group consisting of:

Oct-4, Nanog, and Sox-2.

17. A method of reprogramming a primary somatic cell to a less differentiated state, comprising:

introducing a pluripotency protein into the somatic cell, wherein the pluripotency protein activates expression of an endogenous pluripotency gene, thereby reprogramming the somatic cell to a less differentiated state.

18. The method of claim 17, wherein the pluripotency protein is Oct4, Nanog, or Sox2.

19. The method of claim 17, wherein the endogenous pluripotency gene encodes Oct4 or Nanog.

20. The method of claim 17, wherein the somatic cell does not comprise a selectable marker integrated into the endogenous locus of the pluripotency gene.

21. The method of claim 17, wherein the somatic cell is a human cell or a mouse cell.

22. The method of claim 17, wherein the somatic cell is an adult stem cell.

23. The method of claim 22, wherein the adult stem cell is a hematopoietic stem cell, neural stem cell, or mesenchymal stem cell.