Photoactivatable systems for controlling transcription and ablating synapses.

用于控制转录和消融突触的光激活系统。

基本信息

批准号：
9927247
负责人：
DONALD B ARNOLD
金额：
$ 213.56万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-30 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9927247
关键词：
Ablation Amino Acids Anatomy Binding Binding Proteins Brain Cells Cleaved cell Complex Development Dimerization Excitatory Synapse Exhibits Exposure to Gene Expression Genetic Transcription Goals Grant Halorhodopsins Hour In Vitro Individual Inhibitory Synapse Light Mediating Messenger RNA Morphology Mus Neurons Neurophysiology - biologic function Peptides Process Property Proteins Recombinant Antibody Scaffolding Protein Series Site Slice Specific qualifier value Structure Synapses System Transcriptional Regulation Work Zebrafish base covalent bond experimental study genetic manipulation gephyrin high throughput screening in vivo neural circuit neuronal circuitry new technology novel novel strategies optogenetics photoactivation postsynaptic protein complex recruit tool transcription factor two-photon ubiquitin-protein ligase virtual

项目摘要

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.

用光控制神经元功能的光遗传学工具的出现带来了戏剧性的结果对神经回路的解剖和功能的理解取得了进展。光遗传学工具对于控制转录和改变神经元连接也可能非常有用用于询问神经元回路。然而，这些工具都是基于光激活的配合物，以及此类配合物的所有当前版本，它们依赖于光异构化，在黑暗中具有少量的背景激活，这使得它们很难落实到体内。本次资助的实验使用了一种基于本质上可光裂解的蛋白质 PhoCl，在黑暗中几乎没有表现出激活，因此适合体内使用。我们将利用这个复合体来开发新型光- 用于介导转录和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之前开发的使用 E3 的新技术靶向突触支架蛋白的连接酶介导支架的降解蛋白质。反过来，这会导致突触的结构和功能消融支持。该资助的最终目标是优化 PhoCl 的结构，使其可以被裂解使用两个光子光更快、更高效。