Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Linear and circular long noncoding RNAs regulate cellular reprogramming by sponging multiple miRNAs

ABSTRACT: Using a doxycycline inducible expression of Yamanaka factors (Oct4, Sox2, Klf4 and c-Myc) to generate induced pluripotent stem cells (iPSCs) from mouse embryonic fibroblasts (MEFs), we investigated the dynamic changes of miRNAs, long intergenic noncoding RNAs (lincRNAs) and noncoding circular RNAs (circRNAs) from cells collected at early reprogramming stage. We found that the pools of miRNAs highly expressed in MEFs remain unchanged from day 0 to day 6 after doxycycline induction; unexpectedly, many genes targeted by these miRNAs were actually up-regulated during reprogramming; meanwhile, lincRNAs and circRNAs which have complementary binding sites with the miRNAs were also highly expressed in the reprogramming MEFs (rMEFs). Intriguingly, we found that knockdown of lincRNAs and circRNAs sharing the miRNA binding sites with Oct4 mRNA resulted in down-regulation of Oct4 expression, and significantly decreased the colony formation during reprogramming. Our results suggest that the ceRNA network plays an important role in reprogramming somatic cells to pluripotent stem cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE116684 | GEO | 2019/07/03

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Similar Datasets

RNA-seq profiles of reprogramming cells at Day 3 and Day 6 from MEFs to iPS cells [siNcor1 or siOct4]

Project description:AIM: To detect differences in transcriptional profiles after knocking down Ncor1 or Oct4, compared to a negative control in early reprogramming to pluripotency. DESCRIPTION: RNA-seq profiles of early reprogramming mouse embryonic fibroblasts (MEFs) transduced with lentivirus containing doxycycline-inducible OSKM factors to induce pluripotency . Before starting reprogramming, OSKM-MEFs were transfected with different siRNAs and then they were reprogrammed for 3 or 6 days.

2020-03-27 | GSE139376 | GEO

mRNA expression data from iPSCs, ntESCs and iPSC-nt-ESCs

Project description:We generated three kinds of genetically identical mouse reprogrammed cells: induced pluripotent stem cells (iPSCs), nuclear transfer embryonic stem cells (ntESCs) and iPSC-nt-ESCs that are established after successively reprogramming of iPSCs by nuclear transfer (NT). NtESCs show better developmental potential than iPSCs, whereas iPSC-nt-ESCs display worse developmental potential than iPSCs. We used microarrays to distinguish the gene expression differences among three pluriptoent stem cells and identified that imprinted genes had a similar expression pattern in iPSCs and iPSC-nt-ESCs. We sought to obtain genetic identical pluripotent stem cell lines in order to minimize genetic variations among different reprogrammed cells. To that end, we established a genetically homogenous secondary reprogramming system, in which mouse embryonic fibroblasts (MEFs) carrying doxycycline (dox)-inducible lentiviruses expressing Oct4, Sox2, Klf4 and c-Myc were isolated and used as donors for different reprogramming experiments. We generated iPSCs after plating MEFs in the presence of dox in ES culture conditions, and then derived ntESCs after transplantation of the nucleus from the same MEFs into enucleated oocyte. Furthermore, we successively reprogrammed iPSCs by means of NT and established a set of nt-iPSC lines. Pluripotent stem cells generated from different reprogramming strategies were for RNA extraction and hybridization on Affymetrix microarrays.

2011-05-31 | E-GEOD-24705 | biostudies-arrayexpress

The role of AF10 (Mllt10) in maintaining cell identity (RNA-Seq)

Project description:AF10 is a cofactor of the H3K79 methyltransferase DOT1L. To uncover the role of H3K79me in reprogramming to induced pluripotent stem cells (iPSCs), we bred reprogrammable mice encoding doxycycline inducible Oct4-2A-Klf4-2A-IRES-Sox2-2A-c-Myc (OKSM) transgene with conditional flox (fl)-AF10 (Mllt10). We isolated mouse embryonic fibroblasts (MEFs), deleted AF10 with CRE-recombinase or treated cells with empty vector control, and initiated reprogramming in DOT1L chemical inhibitor SGC0946 or DMSO control. Gene expression was assessed using RNA-Seq and genome wide localization of H3K79me1 and H3K79me2 was determined by ChIP-Seq on day 4 of reprogramming.

2023-04-01 | GSE214139 | GEO

The role of AF10 (Mllt10) in maintaining cell identity (ChIP-Seq)

Project description:AF10 is a cofactor of the H3K79 methyltransferase DOT1L. To uncover the role of H3K79me in reprogramming to induced pluripotent stem cells (iPSCs), we bred reprogrammable mice encoding doxycycline inducible Oct4-2A-Klf4-2A-IRES-Sox2-2A-c-Myc (OKSM) transgene with conditional flox (fl)-AF10 (Mllt10). We isolated mouse embryonic fibroblasts (MEFs), deleted AF10 with CRE-recombinase or treated cells with empty vector control, and initiated reprogramming in DOT1L chemical inhibitor SGC0946 or DMSO control. Gene expression was assessed using RNA-Seq and genome wide localization of H3K79me1, H3K79me2, and RNA Polymerase II (RNAPII) was determined by ChIP-Seq on day 4 of reprogramming.

2023-04-01 | GSE214141 | GEO

Dynamic single-cell imaging of direct reprogramming reveals an early specifying event

Project description:The study of induced pluripotency often relies on experimental approaches that average measurements across a large population of cells, the majority of which do not become pluripotent. Here we used high-resolution, time-lapse imaging to trace the reprogramming process over 2 weeks from single mouse embryonic fibroblasts (MEFs) to pluripotency factor-positive colonies. This enabled us to calculate a normalized cell-of-origin reprogramming efficiency that takes into account only the initial MEFs that respond to form reprogrammed colonies rather than the larger number of final colonies. Furthermore, this retrospective analysis revealed that successfully reprogramming cells undergo a rapid shift in their proliferative rate that corresponds to a reduction in cellular area. This event occurs as early as the first cell division and with similar kinetics in all cells that form induced pluripotent stem (iPS) cell colonies. These data contribute to the theoretical modeling of reprogramming and suggest that certain parts of the reprogramming process follow defined rather than stochastic steps. Gene expression profiling: Mouse E13.5 fibroblasts were generated by blastocyst injection with doxycycline inducible Oct4, Sox2, Klf4, and c-Myc primary iPS cells as previously described. Cells were serum starved with 0.5% FBS containing medium, followed by either continued serum starvation for 96 hours, or by 96 hours of factor induction through medium supplemented with 2 ug/ml doxycycline (Sigma). Two biological replicates of each sample (SerumStarveControl and 96hrInduction) were collected and RNA was isolated in preparation for analysis on Affymetrix Mouse Genome 430 2.0 Arrays as previously described.

2010-04-28 | E-GEOD-21361 | biostudies-arrayexpress

Expression analysis in hESCs with HPAT5-KO

Project description:Long intergenic noncoding RNAs (lincRNAs) are transcribed from thousands of loci in mammalian genomes and play essential roles in diverse biological processes. Their expression is often restricted to distinct developmental contexts and often exhibits precise cell- and/or tissue-specific localization. Recent studies demonstrated that their sequences are highly enriched for repetitive elements (REs) of retroviral origin that might have contributed to their evolution and function-acquisition through development. However, the molecular mechanism(s) by which they regulate gene networks in a cell type-specific manner is largely unknown. Here, we used single-cell gene expression profiling during nuclear reprogramming and identified three endogenous retrovirus 1 (ERV1)-derived lincRNAs (HPAT2, 3 and 5) that are coordinately expressed along with key factors of the core regulatory pluripotency network (OCT4, NANOG, SOX2, SALL4). Chromatin precipitation and reporter-system based assays revealed that NANOG regulates gene expression of these novel lincRNAs in a methylation-dependent manner. Gain- and loss-of function assays identify HPAT5 as a competing endogenous RNA (ceRNA) that acts to regulate pluripotency via a molecular mechanism in which HPAT5 sequesters specific microRNAs (miRNAs) during nuclear reprogramming or human embryonic stem cell (hESC) differentiation to protect SALL4 from let-7 mediated degradation. These results indicate that primate-specific retroviral-derived lincRNAs may regulate widely-conserved pluripotency pathways in human stem cells via novel mechanisms of competition and sequestration. We used microarrays to study the ceRNA crosstalk on a global level in hESCs lacking the HPAT5 locus.

2014-10-06 | E-MTAB-2994 | biostudies-arrayexpress

miR-25 regulates Wwp2 and Fbxw7 and promotes reprogramming of mouse somatic cells

Project description:To reprogram mouse embryonic fibroblasts (MEFs) to induced Pluripotent Stem Cells (iPSCs), we constructed the PiggyBac (PB) transposon carrying the four Yamanaka factor cDNAs controlled by a CAG promoter (PB-CAG-OCKS, Oct4, cMyc, Klf4 and Sox2). As the baseline reprogramming control, we transfected the PB-CAG-OCKS transposon into Oct4-reporter MEFs and plated the cells on STO feeder cells (4F-iPS). To examine the effects of miR-25 on reprogramming, in addition to the Yamanaka factors, we co-transfected the PB-CAG-OCKS plasmid with the PB-CAG-miR-25 plasmid and selected for puromycin resistance (2.0 mg/ml) (25-iPS). We then performed genome-wide gene expression microarray analysis on the iPS cells generated and compared the expression profiles to those of Oct4-reporter MEFs and wildtype ES cells.

2011-10-10 | E-MTAB-409 | biostudies-arrayexpress

ChIP-Seq data for histone marks in mouse embryonic fibroblasts

Project description:The conversion of mouse embryonic fibroblasts (MEFs) to induced pluripotent stem cells (iPS) by forced expression of Oct4, Sox2 and Klf4 is among the earliest demonstrations of reprogramming to a pluripotent state by forced expression of transcription factors. To gain insights into the chromatin state of genes required for reprogramming, we profiled H3K4me3, H3K27me3 and H3K9me3.

2011-01-19 | GSE26657 | GEO

Conversion of murine extraembryonic trophoblast stem cells into induced pluripotent stem cells

Project description:Extraembryonic trophoblast stem cells (TSC) can be converted to induced pluripotent stem cells (TSC-iPSCs) by overexpressing Oct4, Sox2, Klf4 and cMyc. TSC lines were derived from mice harboring a doxycycline inducible Oct4 allele and an Oct4-GFP reporter that has been demonstrated to be activated in cells upon acquisition of pluripotency. Oct4-GFP-positive blastocysts were collected at 3.5 dpc and transduced with lentiviruses encoding doxycycline inducible Sox2, Klf4 and cMyc transgenes (4FTSC). 4FTSC lines were passaged 10 times to establish a population of constantly growing, self-renewing TSCs in the presence of FGF4 and fibroblast conditioned media. To induce lineage conversion, 4FTSCs were cultured under ESC/Lif conditions and doxycycline. After 28 days, several colonies displaying ESC-characteristic dome-shaped colony morphology and bright Oct4-GFP fluorescence could be detected. The 4FTSC-derived colonies were isolated mechanically, dissociated by trypsinization, and plated onto MEFs in ESC medium without doxycycline demonstrating the independence of exogenous factors. They will be called TSC-iPSCs (Trophoblast stem cell derived induced pluripotent stem cells). To examine if the extraembryonic lineage-specific mRNA profile was overcome, the gene-expression profiles of TSC-iPSCs and their parental 4FTSCs were analyzed by microarray analyses and compared to control ESCs.

2011-02-01 | E-GEOD-25872 | biostudies-arrayexpress

Resolving Cell Fate Decisions during Somatic Cell Reprogramming by Single-Cell RNA-seq

Project description:The use of Yamanaka factors or chemical compound converting mouse embryonic fibroblasts (MEFs) into induced pluripotent stem cells (iPSCs). However, these reprogramming processes are highly heterogeneous and remain poorly understood. Here we comprehensively profile the different stages of an optimized Oct4, Sox2, Klf4 and chemical-induced reprogramming at single-cell resolution.

2019-01-01 | GSE103221 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data