Transcription profiling of human ES and keratinocyte cells in response to ionising radiation
Ontology highlight
ABSTRACT: I-131, a source of beta- and gamme radiation and the Auger electron emitter I-125 dU incorporated into nuclear DNA were compared in two cell lines , human ES cells and the keratinocyte cell line HaCaT.
Project description:ES cells are differentiated into mesenderm cell linage by Activin A treatment. During this differentaition, gene expression profile of ES cells, day 4 mesendoderm cell, day 5 mesendoderm cells and mesoderm cells are examined.
Project description:Transcriptional profiling of siRNA-mediated inhibition of CDC7 in IMR90 primary diploid fibroblasts derived from embryonic lung tissue.
Project description:The effect for cell differentiation of phosphorylation of TIF1beta (S824) on the regulation of pluripotency in mouse ES cells was then evaluated. Single amino acid mutagenesis was performed from serine 824 to aspartic acid (SD) or alanine (SA) to mimic phosphorylated or non-phosphorylated versions of TIF1beta, respectively.
Project description:Human embryonic stem cell lines HSF-1 and HSF-6 were differentiated on human fetal gonadal stromal cells for 7 days. The cells were labelled with antibodies against cKIT and SSEA1 and the double positive and double negative cells were collected for microarray analysis.
Project description:Hep3B cells were treated with the chemotherapeutic drug bleomycin. RNA was isolated after 18 h, 24 h and 36 h. The status of the gene expression was examined in response to treatment with bleomycin,
Project description:Epstein-Barr virus (EBV) is able to drive the transformation of B-cells, resulting in the generation of lymphocyte cell lines (LCLs) in vitro. EBV nuclear proteins EBNA3A and EBNA3C are necessary for efficient transformation, while EBNA3B is dispensable. We describe a transcriptome analysis of BL31 cells infected with a series of EBNA3-knockout EBVs, including one deleted for all three EBNA3 genes. BL31 cells were infected with a variety of EBV mutants and their revertants, generated in the EBV-BAC system developed by Henri-Jacques Delecluse and Wolfgang Hammerschmidt. These are mutants of the EBV EBNA3 genes - specifically individual knockouts of EBNA3A, EBNA3B and EBNA3C (in this case the first exon of EBNA3C was retained) and a deletion of the total EBNA3 locus (ie all three EBNA3 genes missing).<br>RNA from cultures of cell lines carrying these mutant EBVs (3-4 independent cell lines for each mutant; 2-3 for each revertant, plus three each for uninfected BL31 and parental B95-8 BAC-infected lines) was isolated using RNeasy midi kits (qiagen) and then ribominus-treated and then labelled and hybridised to Affymetrix Human Exon 1.0ST arrays. Array analysis was performed using the MMBGX algorithm developed by Ernest Turro and Alex Lewin. <br>
Project description:To fully understand the effect of microspheres on genes coding for toxic products, signalling molecules, cell stress responses and so forth. Gene expression profiling using a microarray-based technology allows the expression analysis of thousands of genes simultaneously. This method is more informative than traditional studies on single genes, providing information on networks of gene expression changes in a high-throughput manner. In particular, this approach has been used successfully to study the interaction between cells and their polymers supports yielding enormous information concerning the cell-polymer interactions. Thus, this technique offers a significant contribution towards fully understanding the nature of beadfected cells and the possible transcriptomic alterations that may occur.
Project description:A doxycycline-inducible Xist cDNA transgene was introduced into mouse chromosome 17 at approximately 71.24Mb (NCBI Build m36). Insertion of the transgene was heterozygous, so only one copy of chromosome 17 carried the transgene, and the other chromosome 17 was wild-type. Upon doxycycline induction, the transgene is expressed, producing Xist transgene RNA which localises to autosomal material in cis, leading to silencing of genes adjacent to the transgene. Due to the heterozygous nature of transgene insertion and the cis nature of silencing, for any silenced gene, the maximum possible downregulation in expression levels was 50%.<br><br><br><br>To quantify the changes in the levels of gene expression in response to Xist transgene expression, total RNA was extracted from undifferentiated and differentiated ES cells treated with doxycycline. As background control, total RNA was also prepared from uninduced cells (both undifferentiated and differentiated, cultured in parallel to doxycycline-treated cells). The total RNA was then used to make labelled cRNA, which was hybridised to an Affymetrix GeneChip Mouse Genome 430 2.0 array.