Optogenetic manipulation of cellular communication using engineered myosin motors.
                            
                                blue
                            
                            
                                CRY2olig
                            
                            
                                
                                    Ambystoma mexicanum in vivo
                                
                            
                                
                                    C3H/10T1/2
                                
                            
                                
                                    Cos-7
                                
                            
                            
                                Control of cytoskeleton / cell motility / cell shape
                            
                            
                            
                            
                            
                        
                        
                        
                        
                            Abstract:
                            Cells achieve highly efficient and accurate communication through cellular projections such as neurites and filopodia, yet there is a lack of genetically encoded tools that can selectively manipulate their composition and dynamics. Here, we present a versatile optogenetic toolbox of artificial multi-headed myosin motors that can move bidirectionally within long cellular extensions and allow for the selective transport of GFP-tagged cargo with light. Utilizing these engineered motors, we could transport bulky transmembrane receptors and organelles as well as actin remodellers to control the dynamics of both filopodia and neurites. Using an optimized in vivo imaging scheme, we further demonstrate that, upon limb amputation in axolotls, a complex array of filopodial extensions is formed. We selectively modulated these filopodial extensions and showed that they re-establish a Sonic Hedgehog signalling gradient during regeneration. Considering the ubiquitous existence of actin-based extensions, this toolbox shows the potential to manipulate cellular communication with unprecedented accuracy.