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Determination of the Lef1 regulated tanscriptome in differentiating ES cells

ABSTRACT: Lef1 is a critical transducer of the Wnt/beta-catenin signaling pathway that is essential for mesoderm differentiation in vertebrate embryos. We established a doxycycline inducible ES cell line to over-express Flag-tagged Lef1 in differentiating ES cells which have the capacity to form mesoderm cells in vitro. The goal of this study was to identify the genes/gene networks regulated by Lef1 to understand how it regulates mesoderm differentiation and to establish the Lef1-regulated, Wnt/beta-catenin transcriptome. Total RNA was extracted from ES cells after 3 days of differentiating following Lef1 over-expression from day 2 to 3. We compared the transcriptome of differentially expressed genes in untreated and doxycycline treated ES cells to identify Lef1 regulated genes.

ORGANISM(S): Mus musculus

SUBMITTER: Mark Kennedy

PROVIDER: E-GEOD-73083 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Sp5 and Sp8 recruit β-catenin and Tcf1-Lef1 to select enhancers to activate Wnt target gene transcription.

Kennedy Mark W MW Chalamalasetty Ravindra B RB Thomas Sara S Garriock Robert J RJ Jailwala Parthav P Yamaguchi Terry P TP

Proceedings of the National Academy of Sciences of the United States of America 20160311 13

The ancient, highly conserved, Wnt signaling pathway regulates cell fate in all metazoans. We have previously shown that combined null mutations of the specificity protein (Sp) 1/Klf-like zinc-finger transcription factors Sp5 and Sp8 (i.e., Sp5/8) result in an embryonic phenotype identical to that observed when core components of the Wnt/β-catenin pathway are mutated; however, their role in Wnt signal transduction is unknown. Here, we show in mouse embryos and differentiating embryonic stem cell ...[more]

PMID: 26969725

Similar Datasets

Determination of the Sp5 regulated transcriptome in differentiating ES cells

Project description:Sp5 is a target and critical transducer of the Wnt/beta-catenin signaling pathway and is essential for mesoderm differentiation in vertebrate embryos. We established a doxycycline inducible ES cell line to over-express Flag epitope-tagged Sp5 protein in differentiating ES cells which have the capacity to form mesoderm cells in vitro. The goal of this study was to identify the genes/gene networks regulated by Sp5 to understand how it regulates mesoderm differentiation by identifying important target genes which may mediate its activity. Total RNA was extracted from ES cells after 3 days of differentiating following F-Sp5 over-expression from day 2 to 3. We compared the transcriptome of differentially expressed genes between untreated (minus) and doxycycline treated (plus) ES cells to identify Sp5 regulated genes.

2016-04-01 | E-GEOD-73082 | biostudies-arrayexpress

Genome-wide binding profiles of Sp5 in differentiation mouse embryonic stem cells

Project description:We have determined the genome-wide binding profile of the transcription factor Sp5. Flag-tagged Sp5 was targeted to the HPRT locus in A2Lox.cre ES cells to allow for Doxycycline inducible expression. ES cells were cultured as embryoid bodies for 2 days to prime them for germ layer differentiation. Cells were then treated with Doxycycline for 24 hrs. We found that Flag-Sp5 prefentially bound to promoters associatd with several growth factor signalling pathways, but most prominently with the Wnt/beta-catenin pathway to selectively promote mesoderm differentiation over neural. These data implicate Sp5 as new regulator of Wnt/beta-catenin target expression to promotes ES cell differentation into the mesoderm lineage. Examination of Sp5 transcription factor binding in differentiating embryonic stem cells.

2016-04-01 | E-GEOD-72989 | biostudies-arrayexpress

Sp5 and Sp8 recruit b-catenin to the Tcf1-Lef1 enhanceosome to activate Wnt target gene transcription

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2016-04-01 | E-GEOD-73084 | biostudies-arrayexpress

Determination of the Lef1 regulated tanscriptome in differentiating ES cells

Project description:Lef1 is a critical transducer of the Wnt/beta-catenin signaling pathway that is essential for mesoderm differentiation in vertebrate embryos. We established a doxycycline inducible ES cell line to over-express Flag-tagged Lef1 in differentiating ES cells which have the capacity to form mesoderm cells in vitro. The goal of this study was to identify the genes/gene networks regulated by Lef1 to understand how it regulates mesoderm differentiation and to establish the Lef1-regulated, Wnt/beta-catenin transcriptome.

2016-04-01 | GSE73083 | GEO

Protein and histone binding profiles from mouse ES cells during differentiation and after Kdm6a RNAi treatment.

Project description:Here we show binding and occupancy profiles for KDM6A, H3K27me3 and H3K4me3 to address the epigenetic regulation of a subset of Rhox genes, Rhox6 and 9, in female and male ES cells during differentiation. To further address a functional role for KDM6A in the epigenetic regulation of Rhox6 and 9, binding profiles for female ES cells treated with a control siRNA and siRNA specific for Kdm6a are shown. We report that two members of the Rhox cluster, Rhox6 and 9, are regulated by de-methylation of histone H3 at lysine 27 by KDM6A, a histone demethylase with female-biased expression. Our results are consistent with other homeobox genes in that Rhox6 and 9 are in bivalent domains prior to embryonic stem cell differentiation and thus poised for activation. In female mouse ES cells KDM6A is specifically recruited to Rhox6 and 9 for gene activation, a process inhibited by Kdm6a knockdown. In contrast, KDM6A occupancy at Rhox6 and 9 is low in male ES cells and knockdown has no effect on expression. Our study implicates Kdm6a, a gene that escapes X inactivation, in the regulation of genes important in reproduction, suggesting that KDM6A may play a role in the etiology of developmental and reproduction-related effects of X chromosome anomalies. ChIP-chip was used to analyze the binding profiles of KDM6A, H3K27me3, and H3K4me3 during differentiation in female and male ES cells. Additionally, ChiP-chip of KDM6A binding in control treated and siRNA treated ES cells is presented.

2013-03-22 | E-GEOD-45390 | biostudies-arrayexpress

Extended self-renewal and accelerated reprogramming in the absence of Kdm5b [RNA-Seq]

Project description:ES cell pluripotency is thought to be regulated in part by H3K4 methylation. However, it is unclear how H3K4 demethylation contributes to ES cell function and participates in iPS cell reprogramming. Here, we show that KDM5B, which demethylates H3K4, is important for ES cell differentiation, and presents a barrier to the reprogramming process. Depletion of Kdm5b leads to an extension in the self-renewal of ES cells in the absence of LIF. Transcriptome analysis revealed the persistent expression of pluripotency-genes and underexpression of developmental genes during differentiation in the absence of Kdm5b, suggesting that KDM5B plays a key role in cellular fate changes. We also observed accelerated reprogramming of differentiated cells in the absence of Kdm5b, demonstrating that KDM5B is a barrier to the reprogramming process. Expression analysis revealed that mesenchymal master regulators associated with epithelial-to-mesenchymal transition (EMT) are downregulated during reprogramming in the absence of Kdm5b. Moreover, global analysis of H3K4me3/2 revealed that enhancers of fibroblast genes are rapidly deactivated in the absence of Kdm5b, and genes associated with EMT lose H3K4me3/2 during the early reprogramming process. These findings provide functional insight into the role for KDM5B in regulating ES cell differentiation and as a barrier to the reprogramming process. RNA-Seq of undifferentiated and embryoid body (EB) differentiated murine shLuc and shKdm5b ES cells

2013-10-14 | E-GEOD-47124 | biostudies-arrayexpress

Transcription profiling of mouse embryonic stem cell-derived astrocytes

Project description:Expression profiles for Gfap-positive astrocytes obtained by in vitro differentiation of 129SvJae x C57BL/6 murine embryonic stem (ES) cells. Generated to examine the relationship between expression levels and DNA methylation patterns. Experiment Overall Design: 3 replicates of ES-derived astrocytes.

2008-10-20 | E-GEOD-11483 | biostudies-arrayexpress

Transcription profiling of mouse ES cells, neural precursor cells and embryonic fibroblasts

Project description:Expression profiles for isogenic (129SvJae x C57BL/6) murine embryonic stem (ES) cells, neural precursors (NPC) obtained through in vitro differentiation of the ES cells, and embryonic fibroblasts (MEF) obtained at day 13.5. Experiment Overall Design: 3 replicates of wt ES cells, 3 replicates of wt NPCs obtained by in vitro differentiation, 2 replicated of primary MEFs

2008-06-15 | E-GEOD-8024 | biostudies-arrayexpress

Mouse ES cell differentiated with Activin

Project description:To find miRNAs that involve in renal epithelial differentiation. Mouse ES cells, EB3 (129/Ola) were differentiated with Activin 10 ng/ml after 3 days of embryoid body formation. Cells were harvested on day 18 of the differentiation.

2012-12-05 | E-GEOD-42717 | biostudies-arrayexpress

Investigating the role of Lef1 in mouse embryonic stem cells.

Project description:Genome-wide analysis of gene expression in piggybac empty vector and piggybac-lef1 transfected mouse embryonic stem cells (46C mouse ESCs) Analysis of genes regulated by lef1, and we found that lef1 acts biphasically in mouse embryonic stem cells. Total RNA obtained from empty vector (PB)-transfected 46 ESCs or different states (passage 2 and passage 5) of lef1-transfected (PB-Lef1 P2, PB-Lef1 P5) 46C mouse ESCs.

2016-01-29 | E-GEOD-77330 | biostudies-arrayexpress

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