<em>In Vitro</em> Reconstitution of the Actin Cytoskeleton Inside Giant Unilamellar Vesicles

• DOI: 10.3791/64026-v
• Publication Date: 2022-08-26T00:00:41Z
• Authors: Sheng Chen; Zachary Gao Sun; Michael P. Murrell

The actin cytoskeleton, the principal mechanical machinery in cell, mediates numerous essential physical cellular activities, including cell deformation, division, migration, and adhesion. However, studying dynamics structure of network vivo is complicated by biochemical genetic regulation within live cells. To build a minimal model devoid intracellular regulation, encapsulated inside giant unilamellar vesicles (GUVs, also called liposomes). biomimetic liposomes are cell-sized facilitate quantitative insight into dynamical properties cytoskeleton network, opening viable route for bottom-up synthetic biology. generate encapsulation, inverted emulsion method (also referred to as transfer method) utilized, which one most successful techniques encapsulating complex solutions prepare various cell-mimicking systems. With this method, mixture proteins interest added inner buffer, later emulsified phospholipid-containing mineral oil solution form monolayer lipid droplets. desired generated from droplets crossing lipid/oil-water interface. This enables encapsulation concentrated polymers with components, paving way vitro reconstitution biomimicking network.