Constructs for making induced pluripotent stem cells

Patent metadata

Field	Value
Publication number	US-9228204-B2
Application number	US-201313975004-A
Country	US
Kind code	B2
Filing date	Aug 23, 2013
Priority date	Feb 14, 2011
Publication date	Jan 5, 2016
Grant date	Jan 5, 2016

Abstract

Systems, constructs, and methods for reprogramming cells are provided. In one aspect, for example, a transformation construct for generating iPS cells can include an expression vector having a plurality of reprogramming factors, each reprogramming factor being under control of a separate promoter.

First claim

What is claimed is:

1. A transformation construct for generating induced pluripotent stem (iPS) cells, comprising:
an expression vector including a plurality of reprogramming factors, each reprogramming factor being under control of a separate promoter, wherein the plurality of reprogramming factors includes OCT3/4, SOX2, and at least one member selected from the group consisting of KLF4, c-Myc, NANOG, and LIN28.

2. The construct of claim 1 , wherein the expression vector is selected from the group consisting of plasmids, viruses, and combinations thereof.

3. The construct of claim 1 , wherein the expression vector is selected from the group consisting of adenoviral vectors, episomal vectors, retroviral vectors, and lentiviral vectors.

4. The construct of claim 1 , wherein the expression vector is an episomal vector.

5. The construct of claim 1 , wherein the plurality of reprogramming factors includes OCT3/4, SOX2, KLF4, and c-Myc.

6. The construct of claim 1 , wherein the plurality of reprogramming factors consists of OCT3/4, SOX2, and KLF4.

7. The construct of claim 1 , wherein the plurality of reprogramming factors includes OCT3/4, SOX2, NANOG, and LIN28.

8. The construct of claim 1 , wherein the expression vector has a sequence that is at least 80% homologous to SEQ ID 72.

9. The construct of claim 1 , wherein the expression vector has a sequence that is at least 95% homologous to SEQ ID 72.

10. The construct of claim 1 , wherein at least one of reprogramming factor is under the control of a CMV promoter.

11. The construct of claim 10 , wherein the CMV promoter is a weak CMV promoter.

12. The construct of claim 1 , wherein the expression vector further includes a reporter sequence under control of a separate promoter.

Assignees

• Univ Utah Res Found

Inventors

• Pulst Stefan M
• Paul Sharan
• Dansithong Warunee

Classifications

• C12N2710/10043

  viral genome or elements thereof as genetic vector · CPC title

• C12N5/0619

  Neurons · CPC title

• C12N15/85

  for animal cells · CPC title

• C12N2501/603

  Oct-3/4 · CPC title

• C12N2501/602

  Sox-2 · CPC title