Photoactivatable systems for controlling transcription and ablating synapses.

用于用光控制神经元功能的光遗传学工具的出现已经导致了戏剧性的 对神经回路的解剖和功能的理解的进展。光遗传学工具 用于控制转录和改变神经元连接也是非常有用的 用来询问神经回路然而，这些工具是基于可光活化的 络合物，以及所有现有形式的这种络合物，其依赖于光异构化， 在黑暗中有少量的背景激活，这使得它们很难 在体内实施。本研究中的实验使用了一种新型的光活化复合物， 本质上可光裂解的蛋白质PhoCl，在黑暗中几乎不显示活化， 因此适合于在体内使用。我们将利用这个复合体来开发新的光- 用于介导转录和用于消除兴奋性或抑制性突触的可激活系统。 后一个应用程序基于我们以前开发的使用E3的新技术 靶向突触支架蛋白以介导支架降解的连接酶 proteins.反过来，这导致突触的结构和功能消融， 支持.这项资助的最终目的是优化PhoCl的结构，使其能够被切割。 更快更有效地利用双光子。

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.