Investigating the spatiotemporal regulation of microtubule nucleation in Drosophila. (360G-Wellcome-105653_Z_14_Z)

Microtubules are dynamic polymers that have crucial roles in many eukaryotic cell processes. They are nucleated by multi-protein gamma-TuRCs that are concentrated at various MTOCs within the cell. Although most, if not all, of the gamma-TuRC components are known, it remains unclear how gamma-TuRCs are recruited to different MTOCs at different times during development and during the cell cycle. Moreover, it is not clear how each component functions in the complex. Previous studies have concentrat ed on specific gamma-TuRC components or cell types, and have often studied only mitotic processes. I will use Drosophila as a model to conduct a comprehensive in vivo analysis of gamma-TuRC biology in various cell types, including mitotic cells, developing oocytes and terminally differentiated neurons. Mutant and fluorescently tagged lines exist for most, but not all, of the gamma-TuRC components and I intend to complete this toolset using CRISPR, a novel and rapid genome engineering technique t hat works very efficiently in flies. By studying a range of cell types and examining the role of all gamma-TuRC components, I hope to gain a better understanding of how microtubule nucleation is regulated in space and time throughout animal development.