Methods for reprogramming somatic cells

• US 9,670,464 B2
• Filed: 10/26/2015
• Issued: 06/06/2017
• 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
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US14/923,321
• Publication Number:
  US 20160115456A1
• Time in Patent Office: 589 Days
• 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 composition comprising:

a polynucleic acid encoding an Oct4 protein, and

a polynucleic acid encoding a Sox2 or a Nanog protein;

wherein one or both of the polynucleic acids is operatively linked to at least one heterologous regulatory sequence.

2. The composition of claim 1, further comprising a candidate agent of interest with respect to its potential to reprogram a somatic cell.

3. The composition of claim 2, wherein the candidate agent of interest is a DNA methylation inhibitor, a histone deacetylase inhibitor or PD098059.

4. A composition comprising an isolated somatic cell that comprises an exogenously introduced polynucleic acid encoding Oct4 protein, and an exogenously introduced polynucleic acid encoding Sox2 or Nanog protein.

5. The composition of claim 4, wherein the exogenous polynucleic acids increase Oct4 expression in the isolated somatic cell.

6. The composition of claim 4, wherein the isolated somatic cell is a mammalian cell.

7. The composition of claim 4, wherein the isolated somatic cell is a human cell or a mouse cell.

8. The composition of claim 4, wherein the isolated somatic cell is an adult stem cell.

9. The composition of claim 8, wherein the adult stem cell is selected from the group consisting of:

a hematopoietic stem cell, a neural stem cell, and a mesenchymal stem cell.

10. The composition of claim 4, comprising a candidate agent of interest with respect to its potential to reprogram a somatic cell.

11. The composition of claim 10, wherein the candidate agent of interest is a DNA methylation inhibitor, a histone deacetylase inhibitor or PD098059.

12. A composition comprising one or more recombinant expression cassettes encoding:

a polynucleic acid encoding an Oct4 protein, and a polynucleic acid encoding a Sox2 or a Nanog protein, wherein the polynucleic acids are operably linked to a heterologous promoter.

13. The composition of claim 12, further comprising a candidate agent of interest with respect to its potential to reprogram a somatic cell.

14. The composition of claim 13, wherein the candidate agent of interest is a DNA methylation inhibitor, a histone deacetylase inhibitor or PD098059.