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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Determination of the Sp5 regulated transcriptome in differentiating ES cells

ABSTRACT: 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.

ORGANISM(S): Mus musculus

SUBMITTER: Mark Kennedy

PROVIDER: E-GEOD-73082 | 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

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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

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Determination of the Sp5 regulated transcriptome in differentiating ES cells

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2016-04-01 | GSE73082 | GEO

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

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

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Expression data from differentiating Flk1- and Flk1+ ES cells expressing Snail during Wnt inhibition

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

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