A Chemogenetic Toolkit for Inducible, Cell Type-Specific Actin Disassembly.

Area of Science:

• Cell Biology
• Molecular Biology
• Biochemistry

Background:

• The actin cytoskeleton is crucial for eukaryotic cell functions like morphology, motility, and division.
• Dysregulation of actin is implicated in various diseases.
• Existing tools for studying actin are limited in multicellular contexts.

Purpose of the Study:

• To introduce ChiActs, a next-generation genetic tool for optogenetically perturbing actin nanostructures and dynamics.
• To enable cell type-specific and subcellularly targeted manipulation of actin.
• To investigate the roles of actin nano-assembly in cellular functions.

Main Methods:

• Development of ChiActs based on split DeAct-SpvB components.
• Chemical-induced dimerization or allosteric switching to activate actin disassembly.
• Optogenetic targeting of ChiActs to specific subcellular locations using light.
• Application in various model cell types to assess actin perturbation.

Main Results:

• ChiActs rapidly induce actin disassembly in model cell types.
• ChiActs perturb actin-dependent cellular functions, including lamellipodial protrusions and membrane ruffling.
• ChiActs remodel mitochondrial morphology and reorganize chromatin by localized actin disassembly.

Conclusions:

• ChiActs represent a significant advancement in genetically-encoded tools for actin research.
• These tools facilitate studies on the roles of actin nano-assembly and dynamics in complex biological systems.
• ChiActs unlock new possibilities for investigating actin's involvement in cellular processes and disease.