摘要:SummaryWe present here PhotoGal4, a phytochrome B-based optogenetic switch for fine-tuned spatiotemporal control of gene expression inDrosophilaexplants. This switch integrates the light-dependent interaction between phytochrome B and PIF6 from plants with regulatory elements from the yeast Gal4/UAS system. We found that PhotoGal4 efficiently activates and deactivates gene expression upon red- or far-red-light irradiation, respectively. In addition, this optogenetic tool reacts to different illumination conditions, allowing for fine modulation of the light-dependent response. Importantly, by simply focusing a laser beam, PhotoGal4 induces intricate patterns of expression in a customized manner. For instance, we successfully sketched personalized patterns of GFP fluorescence such as emoji-like shapes or letterform logos inDrosophilaexplants, which illustrates the exquisite precision and versatility of this tool. Hence, we anticipate that PhotoGal4 will expand the powerfulDrosophilatoolbox and will provide a new avenue to investigate intricate and complex problems in biomedical research.Graphical AbstractDisplay OmittedHighlights•PhotoGal4 activates gene expression in response to red light inDrosophilaexplants•Exposure to far-red light allows for PhotoGal4 deactivation•PhotoGal4 can activate gene expression in subgroups of cells within a single tissueOptical Imaging; Genetics; Cell Biology; Technical Aspects of Cell Biology