Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

We describe the reconstitution of dynamic cortical actin networks within cell-like compartments. The approach is based on encapsulation of Xenopus egg extract within water-in-oil emulsions. The growth of cortical actin networks is catalyzed by an amphiphilic actin nucleation-promoting factor that localizes to the water/oil interface. We first describe the preparation of cell-free Xenopus egg extract that provides all the soluble components of the actin machinery. We then describe the preparation of the amphiphilic, fluorescent actin nucleation-promoting factor that directs actin polymerization to the interface. Finally, we describe the steps required for assembly of dynamic actin cortices within water-in-oil emulsions, including the emulsification process and the sample preparation procedures. We provide recommendations for handling sensitive system components and discuss potential uses of this reconstitution approach for cytoskeletal research.

Original publication

DOI

10.1016/bs.mcb.2015.01.011

Type

Journal article

Journal

Methods in cell biology

Publication Date

01/2015

Volume

128

Pages

287 - 301

Addresses

Department of Physics, Technion-Israel Institute of Technology, Haifa, Israel; Russell Berrie Nanotechnology Institute, Technion-Israel Institute of Technology, Haifa, Israel.

Keywords

Ovum, Cell-Free System, Animals, Xenopus, Water, Mineral Oil, Trimethylsilyl Compounds, Actins, Tissue Extracts, Emulsions, Actin Cytoskeleton