A hallmark of profound understanding of the organization of a living cell is the ability to reconstitute essential cellular functionalities from minimal components. To achieve this breakthrough a concerted effort of cell biology, biochemistry and biophysics is required. Our project brings together this expertise to reconstitute the cell’s ability to control the organization of cytoskeletal networks in an artificial ‘Model’ Cell.To achieve a mechanistic understanding of how cell organization is regulated, we will develop methods to manipulate cytoskeletal interactions in space and time and study the effects of such manipulation on functional cytoskeletal organization in the confinement of both artificial systems and cells. We will focus on regulatory interactions at dynamic microtubule plus ends, which play an essential role in cell division, polarization, and migration. Using a combination of in vitro, in vivo, and theoretical approaches, we aim at the following goals:1. Achieve a molecular scale understanding of cooperative and competitive relationships between regulators at microtubule ends, and their effect on microtubule dynamics, microtubule behavior at the cell boundary, and interactions with actin filaments.2. Generate a quantitative understanding of symmetric and polarized positioning of the microtubule cytoskeleton by microtubule-cell boundary interactions during cell division and cell migration.3. Obtain a mechanistic view of microtubule-actin co-organization driven by regulatory effects at microtubule ends, with and without the additional contribution of microtubule-cell boundary interactions, and apply this knowledge to manipulate cell polarization and migration.Synergy between our complementary expertise, tools, infrastructure and local collaboration networks is key to achieving these goals. Our groups are located within short travel distance from each other, allowing the coupling of infrastructure and resources on a daily basis.