Optogenetic control of gene expression in plants in the presence of ambient white light.
                            
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                                EL222
                            
                                PhyB/PIF6
                            
                            
                                
                                    A. thaliana leaf protoplasts
                                
                            
                                
                                    N. benthamiana in vivo
                                
                            
                            
                                Transgene expression
                            
                            
                            
                            
                            
                                Multichromatic
                            
                        
                        
                        
                        
                            Abstract:
                            Optogenetics is the genetic approach for controlling cellular processes with light. It provides spatiotemporal, quantitative and reversible control over biological signaling and metabolic processes, overcoming limitations of chemically inducible systems. However, optogenetics lags in plant research because ambient light required for growth leads to undesired system activation. We solved this issue by developing plant usable light-switch elements (PULSE), an optogenetic tool for reversibly controlling gene expression in plants under ambient light. PULSE combines a blue-light-regulated repressor with a red-light-inducible switch. Gene expression is only activated under red light and remains inactive under white light or in darkness. Supported by a quantitative mathematical model, we characterized PULSE in protoplasts and achieved high induction rates, and we combined it with CRISPR-Cas9-based technologies to target synthetic signaling and developmental pathways. We applied PULSE to control immune responses in plant leaves and generated Arabidopsis transgenic plants. PULSE opens broad experimental avenues in plant research and biotechnology.