Structure guided design of photoselectable channelrhodopsins

光选择性通道视紫红质的结构引导设计

基本信息

批准号：
9244699
负责人：
Vadim Cherezov
金额：
$ 23.1万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-30 至 2018-07-31
项目状态：
已结题

项目摘要

Project Summary: This proposal outlines the development of a fundamentally new optogenetic technology capable of flexibly manipulating the activity of thousands of neurons contributing to the dynamic activity of distributed neural circuits with single neuron resolution. No method that currently exists even remotely meets the need of flexible, selective control of thousands of neurons distributed across large volumes of the brain. Filling this methodological gap is a central research objective of the BRAIN Initiative, because doing so will transform our ability to investigate how the nervous system encodes, processes, utilizes, stores, and retrieves information. The overall objective for this application is to acquire critical structural knowledge of photoactive states of a red-shifted channelrhodopsin and use these to engineer a photoselectable channel prototype that demonstrates the potential of our approach for future development in behaving animals. This would allow opsin-expressing neurons to be flexibly selected, activated, and deselected with light. By leveraging new structural knowledge, we anticipate that we can develop a fundamentally new approach to optogenetics that takes us beyond genetically targeted control and into an era of functionally targeted, flexible control of any neural ensemble. The aims of our research are to obtain the first atomic structures of red-shifted channelrhodopsin mutants in three channel states, engineer a three-state ReaChR mutant with high open conductance and optimized action spectra, and demonstrate reversible photoselective control of neurons in vivo with PReaChR prototypes. We anticipate that completion of these aims will yield the following expected outcomes. First, it will produce new knowledge of the underlying structural transformations between channelrhodopsin photostates that will enable efficient computational design of photoselectable optogenetic tools. Second, it will produce the first examples of photoselective channelrhodopsins useful for neural excitation. Third, it will assess the utility of these new opsins for flexible control of distributed sets of neurons. Collectively, these will provide a roadmap to extending the transformative new trait of photoselectabilty to a wide range of existing optogenetic tools for excitation, inhibition and modulation of neural activity. Further research in this direction should ultimately enable flexible control of spatially complex distributions of neurons in head-fixed and freely moving animals during behavior, a key to furthering our understanding of the intricate neural dynamics that underlie our thoughts, feeling, and actions and how circuit dynamics are disrupted by neurological disorders.

项目概要：该提案概述了一种全新光遗传学技术的开发，该技术能够灵活操纵数千个神经元的活动，从而促进动态活动具有单神经元分辨率的分布式神经电路。目前甚至不存在任何方法远程满足对分布在各处的数千个神经元进行灵活、选择性控制的需求大量的大脑。填补这一方法论空白是该研究的中心研究目标 BRAIN Initiative，因为这样做将改变我们研究神经如何活动的能力系统编码、处理、利用、存储和检索信息。该应用的总体目标是获得光活性的关键结构知识红移通道视紫红质的状态并使用它们来设计光可选择通道原型展示了我们的方法在未来行为发展方面的潜力动物。这将使表达视蛋白的神经元能够被灵活地选择、激活和用光取消选择。通过利用新的结构知识，我们预计我们可以开发一种全新的光遗传学方法，使我们超越遗传学的范畴目标控制并进入了对任何神经系统进行功能目标、灵活控制的时代。我们研究的目的是获得红移通道视紫红质的第一个原子结构三种通道状态的突变体，设计具有高开放性的三态 ReaChR 突变体电导和优化的作用光谱，并证明了可逆光选择性控制具有 PReaChR 原型的体内神经元。我们预计，完成这些目标将产生以下预期成果。首先，它将产生关于之间潜在的结构转变的新知识视紫红质通道蛋白光态将实现光选择的高效计算设计光遗传学工具。其次，它将产生第一个光选择性的例子视紫红质通道对于神经兴奋有用。第三，它将评估这些新的实用性视蛋白用于灵活控制分布式神经元组。总的来说，这些将提供将光选择性的变革性新特性扩展到更广泛的范围的路线图现有的用于激发、抑制和调节神经活动的光遗传学工具。更远这个方向的研究最终应该能够灵活控制空间复杂性头部固定和自由活动的动物在行为过程中神经元的分布，这是加深我们对构成我们思想、感觉、和行为以及神经系统疾病如何扰乱回路动态。