Reprogramming a cell by inducing a pluripotent gene through RNA interference

• US 8,357,666 B2
• Filed: 04/07/2009
• Issued: 01/22/2013
• Est. Priority Date: 08/01/2005
• Status: Expired due to Fees

• Current Assignee: NuPotential, Inc.
• Original Assignee: NuPotential, Inc.
• Inventors: Eilertsen, Kenneth J., Power, Rachel A., Rim, Jong S.
• Primary Examiner(s): Bowman, Amy
• Application Number:
  US12/384,667
• Publication Number:
  US 20090269763A1
• Time in Patent Office: 1,386 Days
• Field of Search: None
• US Class Current: 514/44
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/14 interfering N.A.

Abstract

The invention relate to methods, compositions, and kits for reprogramming a cell. In one embodiment, the invention relates to a method for inducing the expression of at least one gene that contributes to a cell being pluripotent or multipotent. In yet another embodiment, the method comprises inhibiting the expression of a gene that codes for a protein involved in transcriptional repression. In yet another embodiment, the invention relates to a reprogrammed cell or an enriched population of reprogrammed cells that can have characteristics of an ES-like cell, which can be re- or trans-differentiated into a differentiated cell type.

Claims

1. A method for reprogramming a cell comprising:

exposing a cell to an shRNA construct that interferes with expression of a gene that codes for a regulatory protein selected from the group consisting of histone deacetylase and DNA methyltransferase, wherein interfering with expression of said gene induces expression of a pluripotent gene selected from the group consisting of Oct-4, Nanog and Sox-2; and

selecting a reprogrammed cell.

2. The method of claim 1, wherein said selecting said cell comprises:

comparing phenotypes of the cell prior to and after exposure to said shRNA construct, and identifying a cell with a phenotype consistent with a reprogrammed cell.

3. The method of claim 1 further comprising:

expanding the selected cell to a population of cells.

4. The method of claim 1, wherein said selecting a cell comprises:

isolating a cell using an antibody directed to a protein coded for by a pluripotent gene or a cell-surface marker.

5. The method of claim 4, wherein said cell surface marker is selected from the group consisting of:

SSEA3, SSEA4, Tra-1-60, and Tra-1-81.

6. The method of claim 1 further comprising:

prior to selecting said cell, comparing chromatin structure of said pluripotent gene prior to exposure to said shRNA construct to the chromatin structure obtained after exposure to shRNA construct.