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


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2008-08

AUTHORS

Marius Wernig, Christopher J Lengner, Jacob Hanna, Michael A Lodato, Eveline Steine, Ruth Foreman, Judith Staerk, Styliani Markoulaki, Rudolf Jaenisch

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. More... »

PAGES

916-924

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt1483

DOI

http://dx.doi.org/10.1038/nbt1483

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1004160248

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/18594521


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