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Transcriptome analysis of human embryonic stem cells (hESCs) and human fibroblasts upon rapamycin

ABSTRACT: Transcriptome analysis of human embryonic stem cells (hESCs) and human fibroblasts upon rapamycin

PROVIDER: PRJNA422061 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6368100.fastq.gz	Fastqsanger.gz
	SRR6368091.fastq.gz	Fastqsanger.gz
	SRR6368101.fastq.gz	Fastqsanger.gz
	SRR6368092.fastq.gz	Fastqsanger.gz
	SRR6368102.fastq.gz	Fastqsanger.gz

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Transcriptome analysis of human embryonic stem cells (hESCs) and human fibroblasts upon rapamycin

Project description:Rapamycin inhibits mTORC1 complex in a variety of cells. We demonstrated that hESCs are more sensitive to rapamycin treatment as compared to somatic cells (BJ Fibroblasts) and also certain cancer cells (KBM7 cells). To study the molecular mechanisms of this disparity, we performed a transcriptome analysis (RNAseq) of rapamycin-treated hESCs and human fibroblasts for two time points (2 and 4 days of treatment).

2018-04-30 | GSE107965 | GEO

Rapamycin induces chromosome reorganization and increases cytokine production in normal human fibroblasts

Project description:We report the effects of Rapamycin treatment on the transcriptome of normal human dermal fibroblasts isolated from foreskin (designated 2DD). We sequenced mRNA from 2 replicates of proliferative (PRO) quiescent (QUI, serum starved) or treated with 500nM Rapamycin for 5 days (RAP). Comparative analyses with PRO transcripts a baseline indicate that genes that changed expression from Rapamycin treated fibroblasts are significantly different from those of quiescence cells. Rapamycin treated cells showed a significant enrichment for cytokines from the Il-6 cascade.

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Comparison of hiPSCs, hESCs and fibroblasts

Project description:In our study, PRDM14 and NFRkB were found to enhance the reprogramming of human fibroblasts to hiPSCs in the presence of OCT4, SOX2, KLF4, with/without c-MYC (OSK/OSKC). To examnie if the obtained hiPSC share similar expression signature with hESC, we conducted the microarray analysis on the hiPSCs, hESCs and fibroblasts. The result showed that all of the examined hiPSCs resembled hESCs, but differed from fibroblasts MRC5.

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Comparison of hiPSCs, hESCs and fibroblasts

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Expression data from hESCs, hiPSCs and human fibroblasts.

Project description:Detailed analysis comparing hiPSC lines that were newly generated and compared them to already established hiPSC lines We used microarrays to detail various hiPSC lines to confirm faithful reprogramming. hESCs, hiPSCs and human fibroblasts for RNA extraction and hybridization on Affymetrix arrays.

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Mass spectrometry based-proteomic analysis comparing the abundance of RNA-binding proteins (RBPs) between TE03 (I3) human embryonic stem cells (hESCs) and human foreskin fibroblasts (HFFs)

Project description:Here, we used high-resolution mass spectrometry to identify differentially expressed RNA-binding proteins (RBPs) between TE03 (I3) human embryonic stem cells (hESCs) and human foreskin fibroblasts (HFFs). 2102Ep embryonal carcinoma (EC) cells were used as a reference for pluripotent cells. This data accompanies the manuscript: "Uncovering the RNA-binding protein landscape in the pluripotency network of human embryonic stem cells". Abstract: "Embryonic stem cell (ESC) self-renewal and cell-fate decisions are driven by a broad array of molecular signals. While transcriptional regulators have been extensively studied in human ESCs (hESCs), the extent to which RNA-binding proteins (RBPs) contribute to human pluripotency remains unclear. Here, we carry out a proteome-wide screen and identify 810 proteins that directly bind RNA in hESCs. We reveal that RBPs are preferentially expressed in hESCs and dynamically regulated during exit from pluripotency and early lineage specification. Moreover, we show that nearly 200 RBPs are affected by knockdown of OCT4, a master regulator of pluripotency, several dozen of which are directly bound by this factor. Intriguingly, over 20 percent of the proteins detected in our study are putative DNA- and RNA-binding proteins (DRBPs), among them key transcription factors (TFs). Using fluorescently labeled RNA and seCLIP (single-end enhanced crosslinking and immunoprecipitation) experiments, we discover that the pluripotency-associated STAT3 and OCT4 TFs interact with RNA in hESCs and confirm the direct binding of STAT3 to the conserved NORAD long-noncoding RNA. Taken together, our findings indicate that RBPs have a more widespread role in human pluripotency than previously appreciated, reinforcing the importance of post-transcriptional regulation in stem cell biology".

2023-05-05 | PXD025284 | Pride

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Project description:Small RNA molecules, including microRNAs (miRNAs), play critical roles in regulating pluripotency, proliferation and differentiation of embryonic stem cells. miRNA-offset RNAs (moRNAs) are similar in length to miRNAs, align to miRNA precursor (pre-miRNA) loci and are therefore believed to derive from processing of the pre-miRNA hairpin sequence. Recent next generation sequencing (NGS) studies have reported the presence of moRNAs in human neurons and cancer cells and in several tissues in mouse, including pluripotent stem cells. In order to gain additional knowledge about human moRNAs and their putative development-related expression, we applied NGS of small RNAs in human embryonic stem cells (hESCs) and fibroblasts. We found that certain moRNA isoforms are notably expressed in hESCs from loci coding for stem cell-selective or cancer-related miRNA clusters. In contrast, we observed only sparse moRNAs in fibroblasts. Consistent with earlier findings, most of the observed moRNAs derived from conserved loci and their expression did not appear to correlate with the expression of the adjacent miRNAs. We provide here the first report of moRNAs in hESCs, and their expression profile in in comparison to fibroblasts. Moreover, we expand the repertoire of hESC miRNAs. These findings provide an expansion on the known repertoire of small non-coding RNA contents in hESCs.

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Gene expression in human embryonic stem cells (hESCs) upon the differentiation to neural lineages by timed delivery of retinoic acid

Project description:We report gene-expression profiling of hESCs differentiating to neuronal lineages upon the treatment with Retinoic Acid and Smoothened Agonisnt. We study weather RA patterning activity relies on duration of RA exposure and can be used to direct differentiation of hESCs into distinct neuronal cell types.

2020-08-31 | GSE147404 | GEO

A global transcriptome analysis of human dermal fibroblasts upon overexpression of microRNA-132

Project description:MiR-132 is one of the most upregulated miRNAs in human skin wounds at the inflammatory phase of healing; however its biological role in dermal fibroblasts during wound repair has not been studied. To study the genes regulated by miR-132, we transfected miR-132 mimics (pre-miR-132) into primary human dermal fibroblasts to overexpress miR-132. We performed a global transcriptome analysis of fibroblasts upon overexpression of miR-132 using Affymetrix arrays.

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