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Genome-wide analysis of HoxBlinc lncRNA downstream targets in mouse ESC-derived embryonic bodies

ABSTRACT: We find a new non-coding transcript associated with active HoxB locus, which we named HoxBlinc lncRNA. We report that HoxBlinc RNA specifies Flk1+ mesoderm and then promotes the development of HS/PCs and cardiomyocytes. To understand the molecular pathways reguated by HoxBlinc, we generate two doxycycline inducible KD ES R1E cell lines, then differentiate these cell lines to day 6 via embryonic bodies stage. RNA-seq analysis were performed for day 6 EBs with or without doxycycline treatment starting at Flk1+ mesodermal stage.

ORGANISM(S): Mus musculus

PROVIDER: GSE74894 | GEO | 2015/11/11

SECONDARY ACCESSION(S): PRJNA301737

REPOSITORIES: GEO

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Genome-wide analysis of HoxBlinc lncRNA downstream targets in mouse ESC-derived embryonic bodies

Project description:We find a new non-coding transcript associated with active HoxB locus, which we named HoxBlinc lncRNA. We report that HoxBlinc RNA specifies Flk1+ mesoderm and then promotes the development of HS/PCs and cardiomyocytes. To understand the molecular pathways reguated by HoxBlinc, we generate two doxycycline inducible KD ES R1E cell lines, then differentiate these cell lines to day 6 via embryonic bodies stage. RNA-seq analysis were performed for day 6 EBs with or without doxycycline treatment starting at Flk1+ mesodermal stage. RNA from day 6 EBs targeting with 2 invidiual shRNAs with or without doxycycline treatment were combined together for RNA-seq analysis.

2015-11-11 | E-GEOD-74894 | biostudies-arrayexpress

Expression data of control (F/F) and Snai1 knockout (KO) Flk1+/- cells

Project description:Snai1 is a master factor of epithelial to mesenchymal transitioin (EMT), however, its role in embryonic stem cell (ESC) differentiation and lineage commitment remains undefined. We used microarrays to compare the global programme of gene expression between control and Snai1 knockout Flk1+ and Flk1- cells sorted from 4 day EBs. Control and Snai1 knockout ESCs were cultured as embryoid bodies in spotaneous differentiation media, 4 days EBs were dissociated and sorted by anti-Flk1 antibody to separated Flk1+ and Flk1- cells, total RNA were collected for Affymetrix microarrays

2013-12-10 | E-GEOD-53108 | biostudies-arrayexpress

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

Project description:ES cells differentiated in the presence of the Wnt inhibitor DKK1 fail to express the transcription factor Snail and undergo EMT or mesoderm differentiation. We generated an ES cell line, A2.snail, that induced Snail expression upon addition of doxycycline addition. Microarrays were used to gain a global picture of Flk1- and Flk1+ cells generated one day after Snail was expressed during Wnt inhibition. A2.snail ES cells, which express Snail upon addition of doxycycline, were differentiated as embryoid bodies in differentiation media and DKK1. Snail-induced cultures uniquely develop a select population of Flk1+ cells. Total RNA was harvested from sorted control (no doxycycline) Flk1- cells and sorted Snail-induced (doxycycline at day 2) Flk1- and Flk1+ cells at day 3 of differentiation.

2013-10-23 | E-GEOD-26524 | biostudies-arrayexpress

Expression data of control (F/F) and Snai1 knockout (KO) Flk1+/- cells

2013-12-10 | GSE53108 | GEO

Single cell RNA-seq of mouse embryoid bodies and yolk sac

Project description:Blood and endothelial cells arise from hemangiogenic progenitors that are specified from FLK1-expressing mesoderm by the transcription factor ETV2. FLK1 mesoderm also contributes to other tissues, including vascular smooth muscle (VSM) and cardiomyocytes. However, the developmental process of FLK1 mesoderm generation and its derivatives and the lineage relationship among FLK1 mesoderm derivatives these tissues remain obscure. Recent single cell RNA-sequencing (scRNA-seq) studies of early stages of embryogenesis embryos, or in vitro differentiated human embryonic stem (ES) cells have differentiation provided unprecedented information on the spatiotemporal resolution of cells in embryogenesis. Nonetheless, these snapshots still nonetheless offer insufficient information on dynamic developmental processes due to inadvertently missing intermediate states and unavoidable batch effects. Here we performed scRNA-seq of mouse ES cells in asynchronous embryoid bodies (EBs), in vitro differentiated embryonic stem (ES) cells containing undifferentiated ES cells and its differentiated hemangiogenic progeny, as well as yolk sacs, the first hematopoietic extraembryonic tissue in developing embryo that contains hemangiogenic and VSM lineages. We captured a continuous developmental process from undifferentiated pluripotent cells to FLK1 mesoderm-derived tissues involved in hemangiogenesis. This continuous transcriptome map will benefit both basic and applied studies of mesoderm and its derivatives.

2019-11-25 | GSE130146 | GEO

Transcription profiling of mouse embryoid bodies with HOXB4 induced or not induced to identify its target genes

Project description:To unravel the molecular mechanism by which HOXB4 promotes the expansion of early hematopoietic progenitors within differentiating ES cells, we analzed the gene expression profiles of embryoid bodies (EBs) in which transcription of HOXB4 had been induced or not induced. A substantial number of the identified HOXB4 target genes are involved in signaling pathways important for controlling self-renewal, maintenance and differentiation of stem cells. Furthermore, we demonstrate that HOXB4 activity and FGF-signaling are intertwined. HOXB4-mediated expansion of ES cell-derived early progenitors was enhanced by specific and complete inhibition of FGF-receptors. In contrast, the expanding activity of HOXB4 on hematopoietic progenitors in day4-6 embryoid bodies was blunted in the presence of basic FGF (FGF2) indicating a dominant negative effect of FGF-signaling on the earliest hematopoietic cells. Taken together, we show that modulation of FGF signaling is an essential feature of HOXB4 activity in the context of embryonic hematopoiesis. Experiment Overall Design: The Hoxb4i ES cell line (Kyba et al. 2002, Cell 109:29-37) contains an integrated âtet-onâ cassette that allows induction of HOXB4 expression upon treatment with doxycycline. These ES cells can be used to produce hematopoietic cells through the formation of embryoid bodies (EBs). Hematopoiesis starts in these EBs at day 4 and the differentiation into hematopoietic fates can be quantified by colony assays on methyl-cellulose using cells dissociated from EBs at day 6 of incubation. The induction of HOXB4 by incubation with doxycycline increases the production of hematopoietic progenitors within EBs by day 6. Using this specific ES cell line, we compared the transcriptome between embryoid bodies (EBs) in which transcription of HOXB4 had been induced or not induced from day 4 to day 6 (48hours). Experiment Overall Design: Biological replicates: 3

2008-06-16 | E-GEOD-9044 | biostudies-arrayexpress

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

2013-10-23 | GSE26524 | GEO

Transcription profiling by array of human NKX2.1GFP/w spin embryoid bodies treated with all-trans retinoic acid

Project description:The gene targeted NKX2.1GFP/w hESC line was differentiated as spin EBs in BPEL medium supplemented with FGF2 (50-100 ng/ml). At day 7 of the differentiation, the spin EBs were pulsed with all-trans retinoic acid (10-5 M) for 72 hours. At day 10, spin EBs were removed from retinoic acid containing media and transferred from suspension to adherent culture in BEL medium containing no growth factors. Day 23 EBs were FACS sorted based on E-CADHERIN and NKX2.1-GFP expression. Day 0 hESCs, day 7 EBs, day 10 EBs with or without RA treatment and day 23 unsorted EBs and sorted fractions were subjected to Illumina microarray processing.

2011-01-05 | E-MEXP-2807 | biostudies-arrayexpress

Gene expression analysis of murine day 6 embryoid bodies (EBs ) with or without Notch1 (ICN1) induction.

Project description:Analysis of CD41 single positive, VE-cadherin single positive, double positive, and double negatvie populations among 7AAD-CD45- cells from day 6 EBs Doxycline-inducible ICN1 ES cells were differentiated into EBs and ICN1 was induced from day 3-5. Non-induced or induced day 6 EBs were dissociated and subjected to FACS sorting and RNA extraction.

2010-11-14 | E-GEOD-19951 | biostudies-arrayexpress

Transcriptome of Wdr5 and (or) p53 double knockout ESCs and embryoid bodies

Project description:This study describes the transcriptome profiling of day 6 SFEBq embryonic bodies (EBs): 1) WT; 2) Wdr5 KO ; 3) Wdr5 KO with T12h hWDR5 rescue; 4) Wdr5 KO with T48h hWDR5 rescue

2021-06-22 | GSE178555 | GEO

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