Project description:Mouse embryonic stem cells lines derived from different mouse strains show considerable variation concerning their culture conditions, gene targeting efficiency and germ line transmission capacity. To gain insight into the basis for this phenomenon, we generated a SAGE library of the C57BL/6 derived ES cell line Bruce-4 and compared their gene expression to 129 strain-derived cells, which in general show the best performance in gene targeting experiments. Gene Ontology (GO) analyses for functional groups of genes demonstrated considerably higher expression of clusters indicating cellular differentiation as well as cell-cell and cell-matrix adhesion in R1 than in Bruce-4 cells. At the single gene level, almost 90% of the most strongly expressed transcripts in each library were significantly differential. Regarding transcripts that have been linked to stem cell state, seven of the 20 selected genes were expressed in both cell lines, eleven showed stronger expression in Bruce-4 ES cells and only one in R1 cells. These results suggest that expression of “stemness” genes cannot be used as a sole criterion for efficiency of ES cells in gene targeting experiments. Keywords: Embryonic stem cells, SAGE, gene expression profile We generated a SAGE library of the C57BL/6-derived ES cell line Bruce-4 and compared their gene expression to an already available SAGE library of 129-derived R1 ES cells generated by Anisimov et al (GSM72878).
Project description:Mouse embryonic stem cells lines derived from different mouse strains show considerable variation concerning their culture conditions, gene targeting efficiency and germ line transmission capacity. To gain insight into the basis for this phenomenon, we generated a SAGE library of the C57BL/6 derived ES cell line Bruce-4 and compared their gene expression to 129 strain-derived cells, which in general show the best performance in gene targeting experiments. Gene Ontology (GO) analyses for functional groups of genes demonstrated considerably higher expression of clusters indicating cellular differentiation as well as cell-cell and cell-matrix adhesion in R1 than in Bruce-4 cells. At the single gene level, almost 90% of the most strongly expressed transcripts in each library were significantly differential. Regarding transcripts that have been linked to stem cell state, seven of the 20 selected genes were expressed in both cell lines, eleven showed stronger expression in Bruce-4 ES cells and only one in R1 cells. These results suggest that expression of “stemness” genes cannot be used as a sole criterion for efficiency of ES cells in gene targeting experiments. Keywords: Embryonic stem cells, SAGE, gene expression profile
Project description:This sample is a control for the J1, R1 and V6.5 ES samples. The DR4 cell type acts as a feeder cell for stem cells and as such is a small contaminant of the embryonic stem cell growth environment. Keywords: other
Project description:Transcriptional profiling of R1 mouse ES cells infected with non-targeting control (NTC) shRNA and two different shRNA sequences against Cdk8 (shCdk8-1 and shCdk-2) and Med12 (shMed12-1 and shMed12-2).
Project description:To identify whether Cdx2 or Gata3 can activate trophoblast specific gene expression when expressed in R1 ES cells. To assess the dependency of Gata3 activity on Cdx2, Gata3 was also expressed in Cdx2-null ES cells. Keywords: gene expression
Project description:Comparison of trophoblast stem cells vs. R1 ES cells. Experiment Overall Design: this experiment include 2 samples and 14 replicates
Project description:To identify whether Cdx2 or Gata3 can activate trophoblast specific gene expression when expressed in R1 ES cells. To assess the dependency of Gata3 activity on Cdx2, Gata3 was also expressed in Cdx2-null ES cells. Keywords: gene expression a fusion construct of either Cdx2 or Gata3 and the tamoxifen responsive estrogen receptor ligand binding domain was expressed in R1 ES cells. Cell were induced with tamoxifen for 6 days to activate the transcription factor and cultured under trophoblast stem cell conditions