Photoactivatable systems for controlling transcription and ablating synapses.

用于用光控制神经元功能的光遗传工具的出现导致了戏剧性的 了解神经回路的解剖学和功能的进展。光遗传工具 对于控制转录和修改神经元连接也可能是非常有用的 用来审问神经回路。然而，这些工具是基于可照片激活的 络合物，以及依赖于光异构化的此类络合物的所有当前版本， 在黑暗中有少量的背景激活，这使得他们很难 在体内实施。这项资助中的实验使用了一种新型的光激活复合体，基于 这种本质上可光解的蛋白质，PhoCl，在黑暗中几乎没有激活， 因此适合在体内使用。我们将利用这个复合体来开发新的光- 调节转录和消融兴奋性或抑制性突触的可激活系统。 后一种应用基于我们之前开发的使用E3的新技术 以突触支架蛋白为靶点的连接酶介导支架降解 蛋白质。反过来，这会导致突触的结构性和功能性消融 支持。这项拨款的最终目标是优化PhCl2的结构，使其能够被切割 更快、更高效地使用双光子光。

The advent of optogenetic tools for controlling neuronal function with light has led to dramatic advances in the understanding of the anatomy and function of neural circuits. Optogenetic tools for controlling transcription and modifying neuronal connectivity could also be extremely useful for interrogating neuronal circuits. However, these tools are based on photo-activatable complexes, and all current versions of such complexes, which depend on photo-isomerization, have a small amount of background activation in the dark, which makes them difficult to implement in vivo. The experiments in this grant use a novel photo-activation complex based on the intrinsically photo-cleavable protein, PhoCl, which displays virtually no activation in the dark, and thus is appropriate for use in vivo. We will use this complex to develop novel light- activatable systems for mediating transcription and for ablating excitatory or inhibitory synapses. The latter application is based on novel technology that we previously developed that uses E3 ligases targeted to synaptic scaffolding proteins to mediate degradation of the scaffolding proteins. In turn, this results in the structural and functional ablation of synapses that they support. The final aim of the grant is to optimize the structure of PhoCl so that it can be cleaved faster and more efficiently with two photon light.