Single molecule imaging of architectural proteins during zebrafish embryo development

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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.