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

小鼠干细胞及其衍生物染色体域排列的四个维度（空间和时间）分析

• 批准号: 5366406
• 负责人: Professor Dr. Thomas Cremer
• 金额: --
• 依托单位: Walter-Brendel-Zentrum für experimentelle Medizin
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2002
• 资助国家: 德国
• 起止时间: 2001-12-31 至 2006-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5366406?language=en
• 关键词: Analysis; chromosome; territory; arrangements; mouse

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.

在这个项目中，我们比较研究了小鼠胚胎干细胞和从它们分化而来的细胞，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，将分别确定稳定的和动态的安排。我们将应用我们团队中成熟的方法，例如固定细胞核的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，我们还将应用GFP标记的DNA结合蛋白对特定DNA序列进行体内标记。其中一位申请者(SD)在A.Belmont的实验室获得了全面的经验，他是这项技术的先驱。因此，有可能在体内研究上述细胞类型中所述的主题。这个项目将提供对不同细胞类型的高阶染色体区域排列中的差异和相似之处的完整看法的基本见解。