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

在这个项目中，我们比较研究了小鼠胚胎干细胞和由它们分化的细胞（如神经元或巨噬细胞）中染色体区域的结构和排列。通过这种比较，将分别确定稳定和动态安排。我们将应用我们小组中建立的方法，如固定细胞核的3D荧光原位杂交，活细胞核中染色体区域的复制标记，以及3D显微镜和基于计算机的3D和4D（空间加时间）核重建和图像分析。此外，我们将应用在体内标记的特定DNA序列的GFP标记的DNA结合蛋白。申请人之一（SD）在A实验室获得了丰富的经验。贝尔蒙特是这项技术的先驱。因此，可以在所述细胞类型中体内研究所述主题。该项目将提供必要的见解，对不同细胞类型的高阶染色体领土安排的差异和相似性的整体观点。

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.