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

PI: Hochgeschwender, Ute h .提案：1450216标题：BRAIN EAGER：基因编码光源用于非侵入性光遗传学意义光驱动基因靶向神经元激活和沉默的实现为操纵行为实验动物的神经元活动带来了前所未有的可能性。然而，将这种方法转化为临床领域的潜在治疗应用是复杂的，因为需要在大脑中植入光纤作为激活光感视蛋白的光源。该提案描述了一个综合研究，教育和推广计划，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奠定基础。目前还没有其他方法可以实现本文所提出的全范围光子控制神经元的非侵入性。技术描述研究人员将建立在结合光遗传学和生物发光的高度创新的概念上。利用这个概念？因此，他们将利用蛋白质工程技术，通过优化高斯荧光素酶与各种荧光蛋白之间的分子内生物发光共振能量转移（BRET），提高荧光素酶的光输出和延长荧光素酶的发射光谱。他们将在体外测试新结构在激活通道视紫红质和质子和氯化物泵方面的效率。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.