Analysis of chromosome territory arrangements in mouse stem cells and their derivatives in four dimensions (space and time)

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In this project we comparatively investigate the structure and arrangement of chromosome territories in mouse embryonic stem cells and in cells differentiated from them, such as neurons or macrophages. By this comparison, stable and dynamic arrangements will be identified, respectively. We will apply methods well established in our groups, such as 3D fluorescence in situ hybridization of fixed cell nuclei, replication labeling of chromosome territories in nuclei of living cells, as well as 3D microscopy and computer based 3D and 4D (space plus time) nuclear reconstruction and image analysis. In addition, we will apply in vivo labeling of specific DNA sequences by GFP-tagged DNA-binding proteins. One of the applicants (SD) has gained comprehensive experience in the laboratory of A. Belmont, who has pioneered this technique. Thus it becomes possible to investigate the addressed topics in vivo in the mentioned cell types. This project will provide essential insights towards an integral view of differences and similarities in the higher order chromosome territory arrangements of different cell types.