Single molecule imaging of architectural proteins during zebrafish embryo development

细胞核中染色质的组织与基因表达、复制和DNA修复等功能密切相关。在间期，两个重要的组织水平存在于染色体区域内，染色质在活跃区室和非活跃区室中的分离和自相关的染色质环网络。这些环通过控制染色质增强子和启动子区域之间的相互作用参与转录调节。它们通过建筑蛋白及其辅助因子的相互作用动态地产生和溶解。在胚胎早期发育过程中，结构蛋白与染色质的结合是如何介导染色质结构的尚不清楚。在这个项目中，我们想要研究早期胚胎发生中建筑蛋白和染色质之间相互作用的进化。在许多模式生物中，如果蝇和斑马鱼，这个发育时期包含了合子转录的激活。我们计划用单分子荧光成像技术记录斑马鱼胚胎在转录激活期间单个建筑蛋白的动力学行为。因此，我们期望在染色质结构、基因转录和结构蛋白的功能相互作用方面获得新的见解。

The organization of chromatin in the nucleus of a cell is closely interlinked with functions such as gene expression, replication and DNA repair. In interphase, two important levels of organization exist within chromosome territories, separation of chromatin in active and inactive compartments and a network of self-associating chromatin loops. These loops are involved in transcription regulation by controlling the interaction between enhancer and promoter regions of chromatin. They arise and dissolve dynamically through the interplay of architectural proteins and their cofactors. How binding of architectural proteins to chromatin evolves in the course of early embryo development to mediate chromatin architecture is unclear.In this project, we want to study the evolution of interactions between architectural proteins and chromatin during early embryogenesis. In many model organisms such as Drosophila and zebrafish, this developmental period embraces the activation of zygotic transcription. We plan to record the kinetic behaviour of single architectural proteins by single molecule fluorescence imaging in live, developing zebrafish embryos during the time period of transcription activation. Thus we expect to obtain new insights into the functional interplay of chromatin architecture, gene transcription and architectural proteins.