EAPSI: Engineering a Cytoskeleton to Probe the Properties of Cell Membranes

Gutierrez Mary Gertrude L, South San Francisco CA

Investigators

Abstract

The structure and functions of the living cell are highly dependent on the plasma membrane and the cytoskeleton. Understanding the intricate relationship between these two cellular features is challenging and traditional methods only focus on one or the other. In collaboration with Professor Shoji Takeuchi at the University of Tokyo, Japan, this research aims to engineer a cytoskeleton within artificial cells made from model membranes to form a single platform to investigate both cellular features. This collaborative effort will leverage the expertise in direct laser writing (DLW), tissue engineering, and state-of-the-art characterization instruments and techniques at the Takeuchi Laboratory. Cell membranes will be made of giant unilamellar vesicles (GUVs) and will encapsulate DLW structures and will be a cell-free platform allowing for investigating plasma membrane biophysics in response to a cytoskeletal network. The award will exploit the ease of hydrogel-assisted bilayer membrane fabrication to encapsulate a cytoskeletal network made of submicron proteinaceous DLW structures. DLW structures will be formed from protein polymers and bioresponsive materials that will be encapsulated within phospholipid bilayer vesicles in the form of GUVs. This model platform allows investigation on the effects of internal cytoskeletal stress and tension on bilayer membranes and will illuminate changes in membrane properties such as but not limited to shape, permeability, tensegrity response to osmotic gradients, and formation of lipid raft domains due to the cytoskeleton. The successful engineering of a biomimetic cytoskeleton can be extended to investigation of membrane protein functions and further serves as an in vitro artificial cell system to uncover complex cellular interactions. Such investigations will further elucidate crucial cytoskeletal-related function and pathologies such as chromosome separation during mitosis and cellular shape and structuring during intracellular signaling. This award under the East Asia and Pacific Summer Institutes program supports summer research by a U.S. graduate student and is jointly funded by NSF and the Japan Society for the Promotion of Science.