The role of telomeres in 3D organization of the genome within the nucleus
Final Report Abstract
The described project aimed at deciphering the underlying mechanisms of how nuclear organization, specifically the establishment of chromosome territories, is regulated. Previous results had shown that telomeres localize to the nuclear envelope specifically after mitosis, when genome organization is re-established. This led to the hypothesis that telomere anchoring to the nuclear envelope could be involved in guiding chromosomes to their specific locations within the nucleus, which will then remain stable throughout the following interphase. I could establish two very new techniques in the lab, namely APEX proximity biotinylation and CRISPR imaging. Both techniques have the potential to substantially aid in improving our understanding of the functional relevance of human telomere localization throughout the cell cycle. For both techniques, the project was advanced to a point where the protocols are ready to be used to address the questions of 1. how telomeres attach to the nuclear envelope after mitosis (APEX biotinylation to find interaction partners of telomeres with high spatiotemporal specificity), and 2. whether there is a pattern in telomere localization to the nuclear envelope (CRISPR imaging to monitor the movement and relative spatial distribution of specific telomeres). Future studies in the lab utilizing these techniques will aim at answering these questions. Further, the established protocols are ready to be applied to other questions within the lab outside of the scope of this research project. I have started a new postdoc position at the University Hospital in Freiburg, where I will work on gene therapy. Apart from the two techniques described in more detail above, the work at the Salk Institute has taught me a large number of new techniques, including human cell culture, plasmid design, cloning and delivery strategies, and imaging techniques that I will apply in my new project.
