Development of an Optogenetic Tool to Regulate Protein Stability In Vivo
Development of an Optogenetic Tool to Regulate Protein Stability In Vivo
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开发调节体内蛋白质稳定性的光遗传学工具
DOI：
10.1017/9781107281875.011
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通讯作者：
Taxis C
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作者：
Renicke C;Taxis C
引入绿色荧光蛋白作为用于观察活生物体中的生理事件的遗传编码工具彻底改变了生命科学（Tsien，1998; Chudakov等人，2010年）。最近，另一个分支进化出了利用光通过基因表达的光致动器精确操纵细胞功能。旨在观察或调节细胞过程的两种方法都被称为光遗传学（Deisseroth等人，2006; Miesenbock，2009）。第二种策略需要双功能蛋白，其包含用于光接收的传感器结构域和促进特定输出的效应器结构域。光作为一种信号具有某些优势，例如，小分子方法来调节细胞行为或蛋白质活性。这些主要涉及无与伦比的时间和空间控制，以及对光的数量和质量的精确调节。然而，许多生物系统依赖于光作为重要的环境线索并对其作出反应。这在光遗传学方法的实验设计中必须考虑。近年来，已经开发了许多不同的工具，使用光通过调节合成、定位、活性或稳定性来影响蛋白质活性，这已经通过几篇综述进行了深入描述（Gautier等人，2014; Zhang和Cui，2015;齐格勒和Moglich，2015）。这些工具分为两大类：第一类包括天然存在的光致动器，只需要最小的调整，用于异源系统。一个例子是通道视紫红质，它彻底改变了所有水平的神经元研究，从孤立神经元的单细胞测量到整个动物的行为研究（Hausser，2014）。虽然这样的光致动器可以直接转移到选择的实验生物体中，但是通过基于知识的定点诱变或定向进化产生改进的变体是提高这些工具的适用性的一种方式。这为研究人员提供了使用根据其需求优化的实验装置的机会（林，2011）。第二类是合成的模块化光致动器，其通过控制效应器结构域与光感受器的活性来提供新的细胞功能。
The introduction of the green fluorescent protein as a genetically encoded tool for the observation of physiological events in living organisms revolutionized life sciences (Tsien, 1998; Chudakov et al., 2010). Recently, another branch evolved that used light to precisely manipulate cellular functions via genetically expressed photoactuators. Both approaches, aiming at either observation or regulation of cellular processes, are referred to as optogenetics (Deisseroth et al., 2006; Miesenbock, 2009). The second strategy requires a bifunctional protein that comprises a sensor domain for photo-reception and an effector domain facilitating a specific output. Light as a signal has certain advantages compared to, for example, small-molecule approaches to regulate cell behavior or protein activities. Mainly, these relate to unmatched temporal and spatial control, as well as precise regulation of the quantity and quality of light. However, many biological systems rely on and react to light as an important environmental cue. This has to be considered in the experimental design of an optogenetic approach. In recent years, many different tools have been developed using light to influence protein activity by regulating synthesis, localization, activity or stability, which has been described, in depth, by several reviews (Gautier et al., 2014; Zhang and Cui, 2015; Ziegler and Moglich, 2015). These tools fall into two broad classes: the first comprises naturally occurring photoactuators that needed only minimal adjustments for usage in heterologous systems. One example is the channelrhodopsins, which revolutionized neuronal studies at all levels, from single-cell measurements in isolated neurons up to behavioral studies in whole animals (Hausser, 2014). Although such photoactuators might be directly transferable into the experimental organism of choice, the generation of improved variants by knowledge-based, site-directed mutagenesis or directed evolution is a way to improve the applicability of these tools. This offers researchers the opportunity to use an experimental setup that is optimized for their needs (Lin, 2011). The second class is synthetic, modular photoactuators that provide a novel cellular function by controlling the activity of an effector domain with a photoreceptor.