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Unknown,Transcriptomics,Genomics,Proteomics

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Comparative whole-transcriptomic analysis between normal and AKAP-Lbc-depleted human embryonic stem cells

ABSTRACT: Human embryonic stem cells (hESCs) have the unique property of immortality, ability to infinitely self-renew and survive in vitro. In contrast to tumor-deribed cells, their immortality are free from any genomic abberations. Instead, they depend on the AKAP-Lbc/Rho signaling cascade. To understand the downstream way, we performed RNA-seq analyses between normal and AKAP-Lbc-depleted hESCs using the doxycyclin-inducible gene silensing strategy. We use the genetically modified hESCs in which AKAP-13-targeting shRNA is induced by doxycyclin(dox) treatment. To minimize cell loss during treatment, anti-apoptotic factor Bcl-XL is overexpressed. We collected RNA from dox-treated and untreated cells in biological triplicate. We measured gene expression in these 2 sample groups using RNA-seq (illumina HiSeq) .

ORGANISM(S): Homo sapiens

SUBMITTER: Masatoshi Ohgushi

PROVIDER: E-GEOD-67128 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Comparative whole-transcriptomic analysis between normal and AKAP-Lbc-depleted human embryonic stem cells

Project description:Human embryonic stem cells (hESCs) have the unique property of immortality, ability to infinitely self-renew and survive in vitro. In contrast to tumor-deribed cells, their immortality are free from any genomic abberations. Instead, they depend on the AKAP-Lbc/Rho signaling cascade. To understand the downstream way, we performed RNA-seq analyses between normal and AKAP-Lbc-depleted hESCs using the doxycyclin-inducible gene silensing strategy.

2015-08-27 | GSE67128 | GEO

Self-organization of polarized cerebellar plate neuroepithelium in three-dimensional culture of human pluripotent stem cells

Project description:During cerebellar development, the main portion of the cerebellar plate neuroepithelium (NE) gives birth to Purkinje cells and interneurons, while the germinal zone at its dorsal edge, called the rhombic lip (RL), generates granule cells and cerebellar nuclei neurons. However, it remains elusive how these components work together to generate the intricate structure of the cerebellar anlage. In this study, we found that a polarized cerebellar anlage structure self-organizes in three-dimensional (3D) human ES cell (hESC) culture. This NE is capable of differentiating into electrophysiologically functional Purkinje cells. The addition of FGF19 promotes spontaneous generation of dorsoventrally polarized NE structures containing cerebellar and basal plates. Furthermore, further addition of SDF1 promoted the generation of stratified cerebellar plate NE with RL-like germinal zones self-forming at the edge. Thus, hESC-derived cerebellar progenitors exhibit substantial self-organizing potential for generating a polarized structure reminiscent of the early human cerebellar anlage at the first trimester. Examination of mRNA profile in two different treated human ES cells .

2015-04-15 | E-GEOD-63015 | biostudies-arrayexpress

Comparative transcriptomic analysis of self-organized, in vitro generated optic tissues

Project description:Embryonic stem (ES) cells have a remarkable capacity to self-organize complex, multi-layered optic cups in vitro via a culture technique called SFEBq. During both SFEBq and in vivo optic cup development, Rax (Rx) expressing neural retina epithelial (NRE) tissues utilize Fgf and Wnt/β-catenin signalling pathways to differentiate into neural retina (NR) and retinal-pigmented epithelial (RPE) tissues, respectively. How these signaling pathways affect gene expression during optic tissue formation has remained largely unknown, especially at the transcriptome scale. We generated Day 10 Rx+ optic tissue using SFEBq, exposed these tissues to either Fgf or Wnt/β-catenin stimulation, and assayed their gene expression at Days 12 and 15 using RNA-Seq. We measured gene expression in these 5 sample groups in biological triplicate using RNA-seq (Illumina HiSeq) .

2015-05-18 | E-GEOD-62432 | biostudies-arrayexpress

Condensin plays a role in both interphase nuclei and premature chromosome condensation

Project description:Purpose: to identify how condensin I removal affects gene expression globally. Methods: Chicken DT40 CAP-H KO cells were treated with or without dox for 36 h. Total RNA samples were extracted and subjected to sequencing using an Illumina Hiseq2000 platform. Library preparation and Illumina sequencing were performed by Macrogen (South Korea). The sequence tags were spliced-mapped onto the chicken genome galGal4 using Tophat and Bowtie2 following quality test using FASTQC. Differential expression of genes was analyzed using the Bioconductor v2.3 package edgeR v3.2.3. Tag enrichment in NCBI RefSeq genes was calculated between dox-treated and untreated cells using edgeR exact test with tag-wise dispersion estimation. Results: We identified a total of 3798 genes to be differentially expressed in G1 condensin I-depleted chicken DT40 cells: 2495 down regulated and 1303 up regulated. Conclusions: Removal of CAP-H leads to significant misregulation of gene expression, suggesting a key role for condensin I in transcription during interphase. Total RNA profiles of CAP-H KO cells with or without Dox were generated by sequencing using Illumina Hiseq2000 platform.

2017-05-03 | E-GEOD-74892 | biostudies-arrayexpress

MAT2A expression and methionine dependency in Diffuse Midline Gliomas creates a distinct metabolic and epi-genetic sensitivity

Project description:We generated a syngeneic H3K27M diffuse midline glioma (DMG) mouse model and performed total RNA seq on H3K27MPP cells and control cell lines H3wtMPP and Normal Neural stem cells. Aditionally we generate a Dox Inducible Mat2A Knockdown in that human DIPG cell line DIPG04 and performed RNA sequencing on cells treated with 2ug/ml Doxycyclin and no treatment

2022-01-28 | GSE160088 | GEO

ATAC-seq of HEK293 cells with Alu-motif targeted dCas9GFP

Project description:HEK293 cells were selected and subcloned for doxycyclin (DOX) and trimethoprim (TMP) inducible dCas9GFP and a guide RNA molecule targeting an EGA enriched Alu-motif (EEA-g1). Cells were treated with DOX and TMP for three days to induce dCas9GFP expression and ATAC-seq samples were collected from treated and non-treated cells to analyse the effect of targeting the motif with dCas9.

2018-06-07 | E-MTAB-6195 | biostudies-arrayexpress

Transcriptional profile changes by Prox1 expression in thyroid cancer cells

Project description:Human thyroid cancer cell line BCPAP was engineered for doxycyclin-mediated inducible expressoin of Prox1 and next generation sequcing was performed to study the transcriptional regulation by Prox1 BCPAP cells were infected with rTTA2 lentivirus. The resulting cell populations were then stably transfected either with pIRES-hygromycin vector (Clontech) to generate BCPAP-Ctr cells, or with Flag-PROX1-IRES-hygromycin vector to generate BCPAP-Prx cells. PROX1 expression was induced or not in BCPAP-Ctr and BCPAP-Prx cells with 0.5 Âµg/ml doxycycline (Dox) for 48 hours and the total RNA samples were collected for sequencing.

2016-03-02 | E-GEOD-75059 | biostudies-arrayexpress

RNA-seq of hESCs undergoing primed to naive-like resetting via exogenous KLF17 overexpression

Project description:RNA-seq of H9 human embryonic stem cells cultured under naive pluripotency-supportive PXGL conditions either with (Dox) or without (UI) exogenous KLF17 expression for five days. As well as a day 0 control sample (hESCs 24hrs after plating in mTeSR), both UI and +Dox samples were collected at days 1, 2 and 5 after switching to naive-permissive PXGL culture conditions. Only Dox-induced hESCs survive after the first passage and form stable lines in PXGL with naive-like morphology and transcriptome, which could be maintained until naive passage 5 (p5).

2021-09-30 | E-MTAB-10915 | biostudies-arrayexpress

Gene expression comparison of MCF10A cells with and without extra centrosomes through overexpression of PLK4

Project description:The presence of extra centrosomes is a feature of human tumours. However, it is unclear what are the changes elicited by the presence of these abnormalities. To determine the changes in gene expression induced by centrosome amplification, human mammary epithelial cells, MCF10A, expressing a doxycycline inducible PLK4 cDNA were used. Centrosome amplification was induced for 48 hrs upon the addition of doxycyclin to the culture medium. RNA was purified from MCF10A cells without extra centrosomes (no dox) and with extra centrosomes (dox) and processed for microarray analysis.

2018-02-10 | E-MTAB-6415 | biostudies-arrayexpress

Global reprogramming of the cellular translational landscape facilitates cytomegalovirus replication

Project description:Genome-wide profiling establishes that human cytomegalovirus (HCMV) exerts an extensive, unforeseen level of specific control over which cellular mRNAs are recruited to or excluded from polyribosomes. The landscape of translationally-regulated host mRNAs regulates HCMV replication. The HCMV imposed translational signature shares similarities with cancer cells Two biological replicate experiments were performed profiling total and polysomal mRNAs from i) HCMV-infected vs mock-infected cells and ii) uninfected cells transduced with a lentivirus expressing doxycyclin (dox)-inducible HCMV UL38 +/- dox. Analysis of translationally-controlled host genes in HCMV-infected cell and cells expressing the HCMV UL38 gene product

2013-11-01 | E-GEOD-50938 | biostudies-arrayexpress

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