Related Experiment Video
Updated: Jul 16, 2026
A Time-Efficient Fluorescence Spectroscopy-Based Assay for Evaluating Actin Polymerization Status in Rodent and Human Brain Tissues
Published on: June 3, 2021
Design and Use of AsLOV2-Based Optogenetic Tools for Actin Imaging
Kourtney L Kroll1, Tobin R Sosnick2, Ronald S Rock3
1Department of Biochemistry and Molecular Biology & The Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL, USA.
Abstract:
We present protocols for using an optogenetic tool called LILAC for actin imaging. LILAC is a light-controlled version of Lifeact that uses the Avena sativa LOV2 (AsLOV2) domain. By significantly reducing Lifeact's affinity for the cytoskeleton in the dark, LILAC reduces concentration-dependent negative side effects while enabling new image processing methods. We discuss the considerations for using this probe of live-cell actin dynamics, including fluorescent protein selection, cell maintenance, microscopy protocols, and image processing. Our work highlights the potential of AsLOV2-based optogenetics for novel imaging and control tools in cell biology.
