Dataset Information

Gene expression profile in mouse embryonic stem cells in which endogenous Pcl2 expression was stably knocked down using short hairpin RNA

ABSTRACT: Polycomb group (PcG) proteins are highly conserved epigenetic transcriptional repressors important for the control of numerous developmental gene expression programs and have recently been implicated in the modulation of embryonic stem cell (ESC) identity. We identified the PcG protein PCL2 (polycomb-like 2) in a genome-wide screen for novel regulators of self-renewal and pluripotency and predicted that it would play an important role in mouse ESC fate determination. Using multiple biochemical strategies, we provide evidence that PCL2 is a novel Polycomb Repressive Complex 2 (PRC2)-associated protein in mouse ESCs. Knockdown of Pcl2 in ESCs resulted in heightened self-renewal characteristics, defects in differentiation and altered patterns of histone methylation. Through integration of global gene expression and promoter occupancy analyses of both PCL2 and PRC2 components EZH2 and SUZ12, we have predicted PCL2 target genes and formulated regulatory networks describing the role of PCL2 both in modulating transcription of ESC self-renewal genes in undifferentiated ESCs as well as developmental regulators during early commitment and differentiation. Cells were stably expressing Pcl2 shRNA or shRNA mismatch control sequences. Hybridizations of three biological replicates for both the control and Pcl2 shRNA clone were performed.

REANALYSIS of: E-GEOD-16364

ORGANISM(S): Mus musculus

PROVIDER: E-GEOD-16364 | ExpressionAtlas | 2015-09-03

REPOSITORIES: ExpressionAtlas

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Publications

Polycomb-like 2 associates with PRC2 and regulates transcriptional networks during mouse embryonic stem cell self-renewal and differentiation.

Walker Emily E Chang Wing Y WY Hunkapiller Julie J Cagney Gerard G Garcha Kamal K Torchia Joseph J Krogan Nevan J NJ Reiter Jeremy F JF Stanford William L WL

Cell stem cell 20100201 2

Polycomb group (PcG) proteins are conserved epigenetic transcriptional repressors that control numerous developmental gene expression programs and have recently been implicated in modulating embryonic stem cell (ESC) fate. We identified the PcG protein PCL2 (polycomb-like 2) in a genome-wide screen for regulators of self-renewal and pluripotency and predicted that it would play an important role in mouse ESC-fate determination. Using multiple biochemical strategies, we provide evidence that PCL2 ...[more]

PMID: 20144788

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Project description:Embryonic stem cells studies have generated a great interest due to their ability to form a wide variety of matured cells. However, there remains a poor understanding of mechanisms regulating the cell state of ESCs and of the genes they express during early differentiation. Gene expression analysis may be a valuable tool to elucidate either the molecular pathways involved in self-renewal and pluripotency or early differentiation and to identify potential molecular therapy targets. The aim of this study was to characterize at molecular level the undifferentiated mouse ESCs state and the early development towards embryoid bodies. To attemp this issue we performed CodeLink Mouse Uniset I 20K bioarrays in a well-characterized mouse ESC line, MES3, 3- and 7 day-old embryoid bodies and we compared our findings with those in adult tissue cells. Gene expression results were subsequently validated in a commercial stem cell line, CGR8 (ATCC). SAM analysis was used to identify statistically significant changes in microarray data. We identified 3664 genes expressed at significantly greater levels in MES3 stem cells than in adult tissue cells, which included 611 with 3-fold higher gene expression level versus the adult cells. We also investigated the gene expression profile during early embryoid body formation, identifying 2040 and 2243 genes that were up-regulated in 3- and 7-day-old EBs, respectively. Our gene expression results in MES3 cells were partially confirmed in CGR8 cells, showing numerous genes that are expressed in both mouse stem cells. In conclusion, our results suggest that commonly expressed genes may be strong candidates for involvement in maintenance of a pluripotent and undifferentiated phenotype and in early development. This study characterizes at molecular level the undifferentiated mouse ESCs state and the early development towards embryoid bodies. We identified numerous genes not been previously reported, which may offer novel insights into the behaviour of mouse ESC, the maintenance of their undifferentiated state and the mechanisms underlying their pluripotency, self-renewal and early differentiation.