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Prediction and Testing of Novel Transcriptional Networks Regulating Embryonic Stem Cell Self-Renewal and Commitment

ABSTRACT: We have used mouse embryonic stem cells (ESCs) as a model to study the signaling mechanisms that regulate self-renewal and commitment to differentiation. We hypothesized that genes critical to stem cell fate would be dynamically regulated at the initiation of commitment. Time course microarray analysis following initiation of commitment led us to propose a model of ESC maintenance in which highly regulated transcription factors and chromatin remodeling genes (down-regulated in our time course) maintain repression of genes responsible for cell differentiation, morphogenesis and development (up-regulated in our time course). Microarrays of Oct4, Nanog and Sox2 shRNA knockdown cell lines confirmed predicted regulation of target genes. shRNA knockdowns of candidate genes were tested in a novel high throughput screen of self-renewal, confirming their role in ESC pluripotency. We have identified genes that are critical for self-renewal and those that initiate commitment and developed draft transcriptional networks that control self-renewal and early development. Keywords: genetic modification Gene expression in Oct4 knockdown, Sox2 knockdown and their empty vector contol ES cells was analyzed.

ORGANISM(S): Mus musculus

SUBMITTER: Timothy Hughes 

PROVIDER: E-GEOD-7520 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Prediction and testing of novel transcriptional networks regulating embryonic stem cell self-renewal and commitment.

Walker Emily E   Ohishi Minako M   Davey Ryan E RE   Zhang Wen W   Cassar Paul A PA   Tanaka Tetsuya S TS   Der Sandy D SD   Morris Quaid Q   Hughes Timothy R TR   Zandstra Peter W PW   Stanford William L WL  

Cell stem cell 20070601 1

Stem cell fate is governed by the integration of intrinsic and extrinsic positive and negative signals upon inherent transcriptional networks. To identify novel embryonic stem cell (ESC) regulators and assemble transcriptional networks controlling ESC fate, we performed temporal expression microarray analyses of ESCs after the initiation of commitment and integrated these data with known genome-wide transcription factor binding. Effects of forced under- or overexpression of predicted novel regul  ...[more]

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