Nuclear reprogramming factor

• US 8,048,999 B2
• Filed: 12/06/2006
• Issued: 11/01/2011
• Est. Priority Date: 12/13/2005
• Status: Active Grant

• Current Assignee: Kyoto University
• Original Assignee: Kyoto University
• Inventors: Yamanaka, Shinya, Takahashi, Kazutoshi, Okita, Keisuke
• Primary Examiner(s): Crouch; Deborah
• Application Number:
  US12/086,479
• Publication Number:
  US 20090068742A1
• Time in Patent Office: 1,791 Days
• Field of Search: None
• US Class Current: 536/23.5
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/604 Klf-4
• CPC Class Codes: C12N 2501/605 Nanog
• CPC Class Codes: C12N 2501/606 c-Myc
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

There is provided a nuclear reprogramming factor for a somatic cell, which comprises a gene product of each of the following three kinds of genes: an Oct family gene, a Klf family gene, and a Myc family gene, as a means for inducing reprogramming of a differentiated cell to conveniently and highly reproducibly establish an induced pluripotent stem cell having pluripotency and growth ability similar to those of ES cells without using embryo or ES cell.

Claims

1. A nuclear reprogramming factor comprising an isolated Oct family gene, an isolated Klf family gene, and an isolated Myc family gene.

2. The factor according to claim 1, which comprises each of the following three genes:

Oct¾

, Klf4, and c-Myc.

3. The factor according to claim 1, which further comprises an isolated Sox family gene.

4. The factor according to claim 3, wherein the Sox family gene is Sox2 gene.

5. The factor according to claim 1, further comprising a cytokine.

6. The factor according to claim 5, wherein the cytokine is basic fibroblast growth factor (bFGF) and/or stem cell factor (SCF).

7. The factor according to claim 1, which further comprises isolated TERT gene.

8. The factor according to claim 1, which further comprises one or more of the following isolated genes:

SV40 Large T antigen, HPV16 E6, HPV16 E7, and Bmil.

9. The factor according to claim 1, which further comprises one or more of the following isolated genes:

Fbx15, Nanog, ERas, ECAT15-2, Tcl, and β

-catenin.

10. The factor according to claim 1, which further one or more of the following isolated genes:

ECAT1, Esg1, Dnmt3L, ECAT8, Gdf3, Sox15, ECAT15-1, Fthll7, Sall4, Rex1, UTF1, Stella, Stat3, and Grb2.

17. The factor according to claim 2, wherein an L-Myc or N-Myc gene is substituted for the c-Myc gene.

18. The factor according to claim 2, wherein a Klf2 gene is substituted for the Klf4 gene.

19. The factor according to claim 4, wherein a Sox1, Sox3, Sox15 or Sox17 gene is substituted for the Sox2 gene.

20. The factor according to claim 2, which consists of each of the following three genes:

Oct¾

, Klf4, and c-Myc.

21. The factor according to claim 2, further comprising Sox2 gene.

22. The factor according to claim 21, which consists of each of the following four genes:

Oct¾

, Klf4, c-Myc, and Sox2.

23. The factor according to claim 21, wherein an L-Myc or N-Myc gene is substituted for the c-Myc gene.

24. The factor according to claim 23, which consists of each of the following four genes:

Oct¾

, Klf4, L-Myc and Sox2.

11. A nuclear reprogramming factor comprising an isolated Oct family gene, an isolated Klf family gene, and a cytokine.

12. The factor according to claim 11, wherein the cytokine is bFGF and/or SCF.

13. The factor according to claim 11, which comprises each of the following two genes:

Oct ¾ and

Klf4.

14. The factor according to claim 11, which further comprises an isolated Sox family gene.

15. The factor according to claim 14, wherein the Sox family gene is Sox2 gene.

16. The factor according to claim 14, wherein the Oct family gene is an Oct¾

gene, the Klf family gene is a Klf4gene, the Sox family gene is Sox2gene and the cytokine is bFGF.

25. The factor according to claim 16, which consists of Oct¾

gene, Klf4 gene, Sox2 gene and bFGF.