A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types
Graduate School of Biomedical Sciences; Whitehead Institute for Biomedical Research; Department of Cell Biology
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
Life Sciences | Medicine and Health Sciences
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.
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Citation: Nat Biotechnol. 2008 Aug;26(8):916-24. Epub 2008 Jul 1. Link to article on publisher's site
Wernig, Marius; Lengner, Christopher J.; Hanna, Jacob; Lodato, Michael A.; Steine, Eveline; Foreman, Ruth K.; Staerk, Judith; Markoulaki, Styliani; and Jaenisch, Rudolf, "A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types" (2008). GSBS Student Publications. 1531.