A drug-inducible system for direct reprogramming of human somatic cells to pluripotency.
Current approaches to reprogram human somatic cells to pluripotent iPSCs utilize viral transduction of different combinations of transcription factors. These protocols are highly inefficient because only a small fraction of cells carry the appropriate number and stoichiometry of proviral insertions to initiate the reprogramming process. Here we have generated genetically homogeneous "secondary" somatic cells, which carry the reprogramming factors as defined doxycycline (DOX)-inducible transgenes. These cells were obtained by infecting fibroblasts with DOX-inducible lentiviruses, isolating "primary" iPSCs in the presence of the drug, and finally differentiating to "secondary" fibroblasts. When "secondary" fibroblast lines were cultured in the presence of DOX without further viral infection, up to 2% of the cells were reprogrammed to pluripotent "secondary" human iPSCs. This system will facilitate the characterization of the reprogramming process and provides a unique platform for genetic or chemical screens to enhance reprogramming or replace individual factors.
Hockemeyer
Dirk
D
The Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA.
Soldner
Frank
F
The Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA.
Cook
Elizabeth G
EG
The Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA.
Gao
Qing
Q
The Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA.
Mitalipova
Maisam
M
The Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA.
Jaenisch
Rudolf
R
The Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA.|Department of Biology, Massachusetts Institute of Technology, 31 Ames Street, Cambridge, MA 02139 USA.
D054337:Cell Dedifferentiation; D065150:Cellular Reprogramming; D003584:Cytological Techniques; D004318:Doxycycline / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000378:metabolism*
D004318:Doxycycline
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