Tracing and mapping of genome-nuclear lamina interactions in single cells
07 / 2011 - unknown
The 3D architecture of the nucleus is important to organize the genome in compartments of different function. The nuclear lamina (NL), a filamentous protein meshwork underlying the nuclear-membrane, is thought to be important for the spatial organization of chromosomes, and the regulation of gene expression. Genome-wide mapping has revealed that mammalian genomes harbor more than 1,000 large domains that are in molecular contact with the NL. These domains consist mostly of inactive chromatin. While these results have advanced our knowledge of genome organization at the NL considerably, the data were obtained from large asynchronous cell populations. Therefore we still lack insight in the fine structure and dynamics of these interactions in individual cells. I propose to apply a novel tool by which molecular contacts of the genome with the NL can be directly visualized and traced in single living cells. Complementary to this method, I will develop a single-cell genome-wide mapping technique that enables probing the NL interactions of a single entire genome at molecular resolution. Both tools combined will significantly increase our knowledge of the detailed structure and motion of the genome architecture at the NL and the implications of spatial organization on gene regulation in single cells.