GSBS Student Publications

Title

A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types

UMMS Affiliation

Graduate School of Biomedical Sciences; Whitehead Institute for Biomedical Research; Department of Cell Biology

Date

7-3-2008

Document Type

Article

Medical Subject Headings

Animals; *Animals, Genetically Modified; *Cell Dedifferentiation; Chimera; Doxycycline; Epigenesis, Genetic; Fibroblasts; Genetic Vectors; Hybrid Cells; Lentivirus; Mice; Mice, Transgenic; Nuclear Reprogramming; Pluripotent Stem Cells; Transgenes

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

The study of induced pluripotency is complicated by the need for infection with high-titer retroviral vectors, which results in genetically heterogeneous cell populations. We generated genetically homogeneous 'secondary' somatic cells that carry the reprogramming factors as defined doxycycline (dox)-inducible transgenes. These cells were produced by infecting fibroblasts with dox-inducible lentiviruses, reprogramming by dox addition, selecting induced pluripotent stem cells and producing chimeric mice. Cells derived from these chimeras reprogram upon dox exposure without the need for viral infection with efficiencies 25- to 50-fold greater than those observed using direct infection and drug selection for pluripotency marker reactivation. We demonstrate that (i) various induction levels of the reprogramming factors can induce pluripotency, (ii) the duration of transgene activity directly correlates with reprogramming efficiency, (iii) cells from many somatic tissues can be reprogrammed and (iv) different cell types require different induction levels. This system facilitates the characterization of reprogramming and provides a tool for genetic or chemical screens to enhance reprogramming.

Rights and Permissions

Citation: Nat Biotechnol. 2008 Aug;26(8):916-24. Epub 2008 Jul 1. Link to article on publisher's site

DOI of Published Version

10.1038/nbt1483

Related Resources

Link to Article in PubMed

Journal Title

Nature biotechnology

PubMed ID

18594521