Nuclear architecture of gene expression: 3D topology of transcribed and silenced genes in mouse embryonic stem cells and cells differentiated from them

• Applicant: Privatdozent Dr. Steffen Dietzel
• Subject Area: Human Genetics
• Term: from 2003 to 2006
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 5396371

Increasing evidence points to an important role of nuclear organization in regulating the genome. The study applied for will investigate the context of the expression of genes and their localization in the cell nucleus in three dimensions in both, fixed and living cells during differentiation. We will do so by stably transfecting mouse embryonic stem (ES-) cells with transgenes containing a Locus Control Region (LCR) for cell type specific correct regulation plus a tag for a Green Fluorescent Protein (GFP) label. The latter allows microscopic detection of the integrated gene in living and fixed cells. We will observe the transgenes in ES-cells where they are silenced and in cells differentiated from these ES-cells were the transgenes are either transcribed or not transcribed. We will use the b-Globin gene and the chicken lysozyme gene as transgenes. Both are extensively characterized on the molecular level. Thus we will be able to integrate microscopical and molecular observations. Control experiments with the endogenous genes will be performed by multi-color Fluorescence in situ Hybridization (FISH).

• DFG Programme: Research Grants
• Major Instrumentation: Inverted fluorescence microscope
• Instrumentation Group: 5000 Labormikroskope