Engineering Microbial Opsins for Neuroscience via Structure-Guided Recombination

描述(申请人提供)：微生物视蛋白是光敏蛋白质，可通过靶向启动子在特定细胞中表达，并以毫秒的速度打开/关闭，从而提供对细胞功能的遗传和光学(光遗传)控制，具有高度的空间和时间特异性。它们能够控制神经回路的电活动，并提供特定神经元表型的可逆获得和功能丧失，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，这些视蛋白具有广泛的激活光谱，离子选择性有限。 以及狭窄的动力学范围。我们提出了一种利用结构引导的蛋白质工程和定向进化的力量来进行视蛋白工程的新方法。同时，我们还将在利基环境中寻找新的自然产生的视蛋白。扩展的光遗传工具包将有助于研究健康和疾病中的神经元电路。

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.