Structure guided design of photoselectable channelrhodopsins

光选择性通道视紫红质的结构引导设计

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/9244699
• 关键词: Animals; BRAIN initiative; Behavior; Brain; Cell Culture Techniques; Cells; Complex; Crystallography; Data; Development; Disease; Electrophysiology (science); Electrostatics; Engineering; Environment; Feeling; Future; Genetic Markers; Goals; Grant; Head; Health; Knowledge; Light; Lighting; Mammalian Cell; Methods; Motor Cortex; Mus; Mutation; Nervous system structure; Neurons; Neurosciences; Opsin; Optics; Outcome; Pattern; Phase; Process; Property; Protein Engineering; Research; Resolution; Retinal; Roentgen Rays; Shapes; Structure; Synchrotrons; Technology; Thinking; awake; beamline; behavior influence; brain volume; design; flexibility; free-electron laser; in vivo; meetings; mutant; nervous system disorder; neural circuit; neuroregulation; novel; novel strategies; optogenetics; prevent; programs; prototype; relating to nervous system; seal; tool; trait; two-photon

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.

项目摘要： 该提案概述了一种从根本上进行新的光遗传技术的发展 灵活地操纵数千个神经元的活性，导致动态活动 具有单个神经元分辨率的分布式神经回路。 目前没有目前存在的方法 远程满足了对分布在跨越的数千个神经元的灵活，选择性控制的需求 大脑的大量。填补这一方法论差距是 大脑计划，因为这样做将改变我们调查如何紧张的能力 系统编码，过程，利用，存储和检索信息。 该应用程序的总体目的是获得光活性的关键结构知识 红移的频道hopopsin的状态，并使用这些状态来设计可观的频道 原型展示了我们方法对未来发展的潜力 动物。这将使表达Opsin的神经元灵活地，激活和 用光取代。通过利用新的结构知识，我们预计我们可以 开发一种基本的新方法来遗传学，这使我们超越了遗传学 有针对性的控制，并进入功能靶向，灵活地控制任何神经合奏的时代。 我们研究的目的是获得红移通道hopopsin的第一个原子结构 在三个频道状态下的突变体，工程师一个三态触及突变体，高开口 电导和优化的动作光谱，并证明了对 带有原型的体内神经元。 我们预计这些目标的完成将产生以下预期结果。首先，它 将产生有关潜在结构转换之间的新知识 ChannelRhopopsin光静素将实现可观的可观计算设计 光遗传学工具。其次，它将产生照片选择性的第一个示例 ChannelRhopopsins对神经激发有用。第三，它将评估这些新的效用 OPSINS灵活控制分布式神经元集。总的来说，这些将提供 路线图将PhotoleCtabilty的新型新特征扩展到了广泛的范围 现有的光遗传学工具，用于激发，抑制和调节神经活动。更远 朝这个方向进行研究最终应能够灵活控制空间复杂 在行为期间，神经元在头部固定和自由移动动物中的分布，这是一个关键 进一步理解我们思想，感觉， 动作以及电路动力学如何被神经系统疾病破坏。