BRAIN EAGER: Genetically Encoded Light Sources for Non-Invasive Optogenetics

主要研究者：Hochgeschwender，Ute H.提案：1450216标题：BRAIN EAGER：用于非侵入性光遗传学的遗传编码光源的意义光驱动的遗传靶向神经元激活和沉默的实现导致了在行为实验动物中操纵神经元活动的前所未有的可能性。然而，由于需要在大脑中植入光纤作为光源用于激活感光视蛋白。该提案描述了一个综合的研究，教育和推广计划，其重点是开发新一代遗传编码光源，用于光遗传传感器的非侵入性操作。如果成功，这将是一个关键的门槛进步，将为实现微创和高效诊断和治疗的新技术提供基础。目前还没有替代的方法，将实现，非侵入性的，全方位的光子控制的神经元，如这里提出的。技术说明研究人员将建立在高度创新的概念相结合的光遗传学与生物发光。利用这个概念？在临床环境中，他们将利用蛋白质工程来提高光输出，并通过优化高斯荧光素酶和各种荧光蛋白之间的分子内生物发光共振能量转移（BRET）来扩展荧光素酶的发射光谱。他们将测试新结构在体外激活通道视紫红质、质子和氯离子泵的效率。这些概念和试剂的发展将对光遗传学在医学中的应用产生潜在的变革性和广泛的影响。

PI: Hochgeschwender, Ute H.Proposal: 1450216Title: BRAIN EAGER: Genetically Encoded Light Sources for Non-Invasive OptogeneticsSignificanceThe realization of light-driven genetically targeted neuronal activation and silencing has led to unprecedented possibilities in manipulating neuronal activity in the behaving experimental animal.However, translation of this approach into the clinical arena for potential therapeutic applications is complicated by the need for implanting optical fibers in the brain as the light source for activating lightsensing opsins. This proposal describes an integrated research, education, and outreach program which focuses on developing a new generation of genetically encoded light sources for non-invasive manipulation of optogenetic sensors. If successful this will be a key threshold advance that will provide the foundation for new technologies enabling minimally invasive and highly efficient diagnostics and therapies. Currently there are no alternative approaches which would achieve, non-invasively, the full range of photonic control of neurons as proposed here.Technical DescriptionThe investigators will build on the highly innovative concept of combining optogenetics with bioluminescence. To exploit the concept?s potential for non-invasive light activation of optogenetic sensors in clinical settings, they will utilize protein engineering to both improve light output and extend the emission spectrum of the luciferase by optimizing intramolecular bioluminescence resonance energy transfer (BRET) between Gaussia luciferase and various fluorescent proteins. They will test the novel constructs for their efficiency in activating channelrhodopsins and proton and chloride pumps in vitro. The development of these concepts and reagents will have potentially transformative and broad impacts on the implementations of optogenetics in medicine.