A Chemogenetic Toolkit for Inducible, Cell Type‐Specific Actin Disassembly

Abstract The actin cytoskeleton and its nanoscale organization are central to all eukaryotic cells—powering diverse cellular functions including morphology, motility, and cell division—and is dysregulated in multiple diseases. Historically studied largely with purified proteins or in isolated cells, tools to study cell type‐specific roles of actin in multicellular contexts are greatly needed. DeActs are recently created, first‐in‐class genetic tools for perturbing actin nanostructures and dynamics in specific cell types across diverse eukaryotic model organisms. Here, ChiActs are introduced, the next generation of actin‐perturbing genetic tools that can be rapidly activated in cells and optogenetically targeted to distinct subcellular locations using light. ChiActs are composed of split halves of DeAct‐SpvB, whose potent actin disassembly‐promoting activity is restored by chemical‐induced dimerization or allosteric switching. It is shown that ChiActs function to rapidly induce actin disassembly in several model cell types and are able to perturb actin‐dependent nano‐assembly and cellular functions, including inhibiting lamellipodial protrusions and membrane ruffling, remodeling mitochondrial morphology, and reorganizing chromatin by locally constraining actin disassembly to specific subcellular compartments. ChiActs thus expand the toolbox of genetically‐encoded tools for perturbing actin in living cells, unlocking studies of the many roles of actin nano‐assembly and dynamics in complex multicellular systems.