Reprogramming of murine fibroblasts to induced pluripotent stem cells with chemical complementation of Klf4.
Ectopic expression of defined transcription factors can reprogram somatic cells to induced pluripotent stem (iPS) cells, but the utility of iPS cells is hampered by the use of viral delivery systems. Small molecules offer an alternative to replace virally transduced transcription factors with chemical signaling cues responsible for reprogramming. In this report we describe a small-molecule screening platform applied to identify compounds that functionally replace the reprogramming factor Klf4. A series of small-molecule scaffolds were identified that activate Nanog expression in mouse fibroblasts transduced with a subset of reprogramming factors lacking Klf4. Application of one such molecule, kenpaullone, in lieu of Klf4 gave rise to iPS cells that are indistinguishable from murine embryonic stem cells. This experimental platform can be used to screen large chemical libraries in search of novel compounds to replace the reprogramming factors that induce pluripotency. Ultimately, such compounds may provide mechanistic insight into the reprogramming process.
Lyssiotis
Costas A
CA
Department of Chemistry, Scripps Research Institute, La Jolla, CA 92037, USA.
Foreman
Ruth K
RK
Staerk
Judith
J
Garcia
Michael
M
Mathur
Divya
D
Markoulaki
Styliani
S
Hanna
Jacob
J
Lairson
Luke L
LL
Charette
Bradley D
BD
Bouchez
Laure C
LC
Bollong
Michael
M
Kunick
Conrad
C
Brinker
Achim
A
Cho
Charles Y
CY
Schultz
Peter G
PG
Jaenisch
Rudolf
R
D000818:Animals; D001552:Benzazepines / Q000494:pharmacology*; D002454:Cell Differentiation; D044127:Epigenesis, Genetic / Q000187:drug effects*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D017930:Genes, Reporter; D018398:Homeodomain Proteins / Q000235:genetics; D007211:Indoles / Q000494:pharmacology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D051379:Mice; D000071317:Nanog Homeobox Protein; D039904:Pluripotent Stem Cells / Q000166:cytology*; D054852:Small Molecule Libraries / Q000494:pharmacology*
D001552:Benzazepines; D018398:Homeodomain Proteins; D007211:Indoles; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D054852:Small Molecule Libraries; C119620:kenpaullone; D008156:Luciferases