de Mena Lorena, Rincon-Limas Diego E
Department of Neurology, McKnight Brain Institute, and Norman Fixel Institute for Neurological Diseases, University of Florida, Gainesville, FL 32611, USA.
Department of Neurology, McKnight Brain Institute, and Norman Fixel Institute for Neurological Diseases, University of Florida, Gainesville, FL 32611, USA; Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of Florida, Gainesville, FL 32611, USA; Genetics Institute, University of Florida, Gainesville, FL 32611, USA.
iScience. 2020 Jul 24;23(7):101308. doi: 10.1016/j.isci.2020.101308. Epub 2020 Jun 23.
We present here PhotoGal4, a phytochrome B-based optogenetic switch for fine-tuned spatiotemporal control of gene expression in Drosophila explants. 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 in Drosophila explants, which illustrates the exquisite precision and versatility of this tool. Hence, we anticipate that PhotoGal4 will expand the powerful Drosophila toolbox and will provide a new avenue to investigate intricate and complex problems in biomedical research.
我们在此展示光激活Gal4（PhotoGal4），这是一种基于光敏色素B的光遗传学开关，用于对果蝇外植体中的基因表达进行精细的时空控制。该开关整合了植物中光敏色素B与PIF6之间的光依赖性相互作用以及酵母Gal4/UAS系统的调控元件。我们发现，PhotoGal4分别在红光或远红光照射下能有效激活和失活基因表达。此外，这种光遗传学工具对不同的光照条件有反应，从而实现对光依赖性反应的精细调节。重要的是，通过简单地聚焦激光束，PhotoGal4能以定制的方式诱导复杂的表达模式。例如，我们成功地在果蝇外植体中勾勒出了绿色荧光蛋白（GFP）荧光的个性化图案，如表情符号形状或字母标志，这说明了该工具的精准度和多功能性。因此，我们预计PhotoGal4将扩展强大的果蝇工具库，并为研究生物医学研究中复杂的问题提供一条新途径。
