Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Gene expression profile after Dax1 deletion in ES cells

ABSTRACT: To investigate the function of Dax1 in mouse ES cells, whole genome microarray expression profiling was employed to identify genes that are potentially affected by the deletion of Dax1. An inducible knockout ES cell line of Dax1 with the Cre-loxP system was generated and gene expression was measured (i) without Dax1 deletion, (ii) after 4 days of Dax1 deletion, or (iii) after several passags of Dax1 deletion. Each sample was prepared in triplicate.

ORGANISM(S): Mus musculus

SUBMITTER: Hitoshi Niwa

PROVIDER: E-GEOD-61152 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Nr0b1 is a negative regulator of Zscan4c in mouse embryonic stem cells.

Fujii Setsuko S Nishikawa-Torikai Satomi S Futatsugi Yoko Y Toyooka Yayoi Y Yamane Mariko M Ohtsuka Satoshi S Niwa Hitoshi H

Scientific reports 20150316

Nuclear receptor subfamily 0, group B, member 1 (Nr0b1, also known as Dax1) is regarded as an important component of the transcription factor network that governs pluripotency in mouse embryonic stem (ES) cells. Here we generated inducible knockout ES cells for Nr0b1 using the Cre-loxP system and analyzed its precise function. We succeeded in establishing the Nr0b1-null ES cells and confirmed their pluripotency by showing their contribution to chimeric embryos. However, they proliferated slowly ...[more]

PMID: 25772165

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Project description:The molecular hallmark of the Ewing family of tumors is the presence of balanced chromosomal translocations leading to the formation of chimerical transcription factors (i.e. EWS/FLI1) that play a pivotal role in the pathogenesis of Ewing tumors by deregulating gene expression. We have recently demonstrated that DAX1 (NR0B1), an orphan nuclear receptor which was not previously implicated in cancer, is induced by the EWS/FLI1 oncoprotein and is highly expressed in Ewing tumors, suggesting that DAX1 is a biologically relevant target of EWS/FLI1-mediated oncogenesis. In this work we demonstrate that DAX1 is a direct transcriptional target of the EWS/FLI1 oncoprotein through its binding to a GGA-rich region in the DAX1 promoter and show that DAX1 is a key player of EWS/FLI1-mediated oncogenesis. DAX1 silencing using an inducible model of RNA interference induces growth arrest in the A673 Ewing cell line and severely impairs its capability to grow in semisolid medium and form tumors in immunodeficient mice. Gene expression profile analysis demonstrated that about ten percent of the genes regulated by EWS/FLI1 in Ewing cells are DAX1 targets, confirming the importance of DAX1 in Ewing oncogenesis. These findings indicate that DAX1 is an important player in the pathogenesis of the Ewing family of tumors, identify new functions for DAX1 as a cell cycle progression regulator and open the possibility to new therapeutic approaches based on DAX1 function interference. A673 cells derived from Ewing sarcoma were genetically enginereed to express specific shRNAs against GFP (control), EWS/FLI1 and DAX1 upon doxycycline stimulation. Three independent clones and a polyclonal population from each enginereed cell were analyzed. Cells were stimulated with doxycycline for 72 hours to induce the expression of the corresponding shRNA and whole gene expression profile performed. Gene expression profile in A673 cells in which EWS/FLI1 or DAX1 were silenced were compared to the control cells.

Dataset Information

Gene expression profile after Dax1 deletion in ES cells

Publications

Nr0b1 is a negative regulator of Zscan4c in mouse embryonic stem cells.

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