title: A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types.
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.
authors.affiliation: Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, Massachusetts 02142, USA.
authors.forename: Marius
authors.initials: M
authors.lastname: Wernig
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lengner
authors.forename: Jacob
authors.initials: J
authors.lastname: Hanna
authors.forename: Michael A
authors.initials: MA
authors.lastname: Lodato
authors.forename: Eveline
authors.initials: E
authors.lastname: Steine
authors.forename: Ruth
authors.initials: R
authors.lastname: Foreman
authors.forename: Judith
authors.initials: J
authors.lastname: Staerk
authors.forename: Styliani
authors.initials: S
authors.lastname: Markoulaki
authors.forename: Rudolf
authors.initials: R
authors.lastname: Jaenisch
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D054337:Cell Dedifferentiation; D065150:Cellular Reprogramming / Q000187:drug effects*; D002678:Chimera / Q000235:genetics; D004318:Doxycycline / Q000494:pharmacology*; D044127:Epigenesis, Genetic; D005347:Fibroblasts / Q000166:cytology; D005822:Genetic Vectors; D006822:Hybrid Cells; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects*; D019076:Transgenes
chemical_list: D004318:Doxycycline