Second generation genetically encoded photosensitizers and killers for systems neuroscience

直到今天，光遗传学试剂的新发展仍在不断改变神经科学。在这个过程中，已经设计了旨在体内细胞的有效光消融的工具。这些试剂中的一些被证明对于某些体内应用是有用的。然而，似乎在遗传编码的发色团的帮助下光遗传学消融细胞尚未成为回路神经科学中的通用工具，并且该方法需要改进以实现其全部潜力。我们在这里提出了一个合作的努力，旨在工程高灵敏度的第二代光遗传学工具的可靠的光消融的遗传靶向细胞在体内。Griesbeck和雷夫的实验室基于两个在光照下产生活性氧（ROS）的蛋白质家族，提议联合工程（Griesbeck）和体内应用/验证（雷夫）具有改进特性的新的ROS产生蛋白质。 候选人将在体内果蝇的生理和行为实验中进行定量测试。因此，我们将集中在视觉处理的实验，包括处理的光流，视觉控制的导航和神经元计算的颜色视觉。 改进的蛋白质应该包括经典光遗传学试剂的强大特征，如具有高空间分辨率和细胞特异性的激活，应该在亚细胞水平上工作以探测回路内的棘或突触的作用，并且在很大程度上与现有的光遗传学报告基因正交。因此，用于光诱导消融的新的ROS生成蛋白质可能对于遗传模型生物的神经科学研究非常有用，而且对于生物学和生物医学中的许多其他应用也非常有用。

New developments of optogenetic reagents are constantly transforming neuroscience until today. In the course of this process, tools have been engineered that aim at efficient photo-ablation of cells in vivo. Some of these reagents demonstrated usefulness for certain applications in vivo. However, it appears that optogenetic ablation of cells with the aid of genetically encoded chromophores has not yet become a versatile tool in circuit neuroscience and that the method requires improvement to realize its full potential. We here propose a collaborative effort that aims at engineering highly sensitive second generation optogenetic tools for the reliable photo-ablation of genetically targeted cells in vivo. Building on two families of proteins that produce reactive oxygen species (ROS) upon illumination, the labs of Griesbeck and Reiff propose to join forces for the engineering (Griesbeck) and in vivo application/verification (Reiff) of new ROS-producing proteins with improved properties. Candidates will be quantitatively tested in physiological and behavioral experiments in Drosophila melanogaster in vivo. Thereby, we will focus on experiments on visual processing including the processing of optic flow, the visual control of navigation and neuronal computations underlying color vision. Improved proteins should include powerful features of classical optogenetic reagents like activation with high spatial resolution and cellular specificity, should as work on the subcellular level to probe the roles of spines or synapses within a circuit, and be largely orthogonal to existing optogenetic reporters. Thus, new ROS-generating proteins for light-induced ablation may become highly useful for neuroscience research in genetic model organisms, but also for a number of other applications in biology and biomedicine.