Design and Use of AsLOV2-Based Optogenetic Tools for Actin Imaging.

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.