Dissecting self-renewal in stem cells with RNA interference.
We present an integrated approach to identify genetic mechanisms that control self-renewal in mouse embryonic stem cells. We use short hairpin RNA (shRNA) loss-of-function techniques to downregulate a set of gene products whose expression patterns suggest self-renewal regulatory functions. We focus on transcriptional regulators and identify seven genes for which shRNA-mediated depletion negatively affects self-renewal, including four genes with previously unrecognized roles in self-renewal. Perturbations of these gene products are combined with dynamic, global analyses of gene expression. Our studies suggest specific biological roles for these molecules and reveal the complexity of cell fate regulation in embryonic stem cells.
Ivanova
Natalia
N
Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA. nivanova@molbio.princeton.edu
Dobrin
Radu
R
Lu
Rong
R
Kotenko
Iulia
I
Levorse
John
J
DeCoste
Christina
C
Schafer
Xenia
X
Lun
Yi
Y
Lemischka
Ihor R
IR
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation; D004268:DNA-Binding Proteins / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology; D015870:Gene Expression; D005816:Genetic Complementation Test; D018398:Homeodomain Proteins / Q000378:metabolism; D051379:Mice; D000071317:Nanog Homeobox Protein; D034622:RNA Interference; D012038:Regeneration / Q000235:genetics* / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse