Engineering Microbial Opsins for Neuroscience via Structure-Guided Recombination

描述（由申请人提供）：微生物视蛋白是一种光敏蛋白，可以通过靶向启动子在特定细胞中表达，并以毫秒速度打开/关闭，从而提供具有高空间和时间特异性的细胞功能的遗传和光学（“光遗传”）控制。它们能够控制神经回路的电活动，并揭示特定神经元表型功能的可逆增益和丧失，这使我们能够以前所未有的方式研究神经系统和疾病。然而，今天的光遗传学研究依赖于一组有限的天然微生物视蛋白，它们具有广泛的激活光谱，有限的离子选择性

DESCRIPTION (provided by applicant): Microbial opsins are light-sensitive proteins that can be expressed in specified cells via targeting promoters and turned on/off with millisecond speed, thus providing genetic and optical ('optogenetic') control of cell function with high spatial and temporal specificity. Their ability to control the electrical activity of neural circuits and confe reversible gain and loss of function of specific neuronal phenotypes allows us to study neural systems and diseases in unprecedented manner. Optogenetic research today, however, relies on a limited set of natural microbial opsins with broad activation spectra, limited ion selectivity and a narrow range of kinetics. We are proposing a novel approach to opsin engineering that capitalizes on the power of structure-guided protein engineering and directed evolution. In parallel, we will also search for novel naturally occurring opsins in niche environments. The expanded optogenetic toolkit will facilitate the investigation of neuronal circuits in health and disease.