title: Building an artificial actin cortex on microscopic pillar arrays.
abstract: Eukaryotic cells obtain their morphology and mechanical strength from the cytoskeleton and in particular from the cross-linked actin network that branches throughout the whole cell. This actin cortex lies like a quasi-two-dimensional (2D) biopolymer network just below the cell membrane, to which it is attached. In the quest for building an artificial cell, one needs to make a biomimetic model of the actin cortex and combine this in a bottom-up approach with other "synthetic" components. Here, we describe a reconstitution method for such an artificial actin cortex, which is freely suspended on top of a regular array of pillars. By this immobilization method, the actin network is only attached to a surface at discrete points and can fluctuate freely in between. By discussing the method to make the micropillars and the way to reconstitute a quasi-2D actin network on top, we show how one can study an isolated, reconstituted part of a cell. This allows the study of fundamental interaction mechanisms of actin networks, providing handles to design a functional actin cortex in an artificial cell.
authors.affiliation: Natuur- en Sterrenkunde and LaserLab, Vrije Universiteit, De Boelelaan, Amsterdam, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: Ayadi
authors.affiliation: Natuur- en Sterrenkunde and LaserLab, Vrije Universiteit, De Boelelaan, Amsterdam, The Netherlands.
authors.forename: W H
authors.initials: WH
authors.lastname: Roos
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001704:Biopolymers / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology*; D008870:Microtubules / Q000378:metabolism; D011817:Rabbits
chemical_list: D000199:Actins; D001704:Biopolymers
keywords: Artificial cell; Biophysics; Biopolymer; Cross-linked network; Cytoskeleton; Fluorescence microscopy; In vitro reconstitution; Microstructured surfaces; PDMS; Synthetic biology