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Transcriptomes of miR-290-295 KO mESCs treated with siPOOLs against the transcription factor Tfap4

ABSTRACT: Transcriptomes of miR-290-295 KO mESCs treated with siPOOLs against the transcription factor Tfap4

PROVIDER: PRJNA751876 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15338351.fastq.gz	Fastqsanger.gz
	SRR15338352.fastq.gz	Fastqsanger.gz
	SRR15338353.fastq.gz	Fastqsanger.gz
	SRR15338354.fastq.gz	Fastqsanger.gz
	SRR15338355.fastq.gz	Fastqsanger.gz

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Transcriptomes of miR-290-295 KO mESCs treated with siPOOLs against the transcription factor Tfap4

Project description:In order to identify direct microRNA targets in mouse embryonic stem cells (mESCs), we knocked out microRNA effector genes, profiled their transcriptomes and integrated the data with additional published OMICs data sets. To validate the analysis, we generated miR-290-295 KO mutant cell lines, assessed the transcriptome profile and compared the observed misregulation to the predictions of the analysis. We identified the transcription factor Tfap4 as novel potential direct and functional target of miR-290-295. To assess the role of Tfap4, we rescued its expression in the miR-290-295 KO cell line using siPOOLs and profiled the transcriptome.

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Project description:In order to identify direct microRNA targets in mouse embryonic stem cells (mESCs), we generated miR-290-295 KO mutant cell lines, assessed their transcriptomes. We identified novel transcription factors (TF) as potential direct and functional targets of miR-290-295. To assess the role of these TFs, we rescued their expression in the miR-290-295 KO cell line using siPOOLs and profiled their transcriptomes.

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Project description:MicroRNAs (miRNAs) post-transcriptionally regulate the expression of thousands of distinct mRNAs. While some regulatory interactions help to maintain basal cellular functions, others are likely relevant in more specific settings, such as response to stress. Here we describe such a role for the mir-290-295 cluster, the dominant miRNA cluster in mouse embryonic stem cells (mESCs). Examination of a target list generated from bioinformatic prediction, as well as expression data following miRNA loss, revealed strong enrichment for apoptotic regulators, two of which we validated directly: Caspase 2, the most highly conserved mammalian caspase, and Ei24, a p53 transcriptional target. Consistent with these predictions, mESCs lacking miRNAs were more likely to initiate apoptosis following genotoxic exposure to gamma irradiation or doxorubicin. Knockdown of either candidate partially rescued this pro-apoptotic phenotype, as did transfection of members of the mir-290-295 cluster. These findings were recapitulated in a specific mir-290-295 deletion line, confirming that they reflect miRNA functions at physiological levels. In contrast to the basal regulatory roles previously identified, the pro-survival phenotype shown here may be most relevant to stressful gestations, where pro-oxidant metabolic states induce DNA damage. Similarly, this cluster may mediate chemotherapeutic resistance in a neoplastic context, making it a useful clinical target.

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