Engineering of Chrimson for Subcellular Optogenetic application

用于亚细胞光遗传学应用的 Chrimson 工程

• 批准号: 425994138
• 负责人: Professor Dr. Marcus Elstner
• 金额: --
• 依托单位: Abteilung für Theoretische Chemische Biologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/425994138?language=en
• 关键词: Engineering; Chrimson; Subcellular; Optogenetic; application

The aim of the project is to further characterize and engineer Chrimson, the channel-rhodopsin with the most red shifted absorption spectrum and high proton selectivity, which shares structural features both with archaeal light driven proton pumps and most other channelrhodopsin. We will focus our studies on the unusual substructures including the active site, the outer gate, and the selectivity filter by using biochemical, biophysical and theoretical approaches. For a thorough understanding, we will build and investigate computational models, which will help to interpret experimental findings and assist in the rational design of red-shifted variants. We will engineer new and even more red-shifted variants with increased sodium-selectivity and/or modified kinetics. Due to the increased transmission of neuronal tissues at wavelength beyond 600 nm, these Chrimson derivatives are of outmost interest for in vivo applications in neuroscience. Furthermore, we will explore the biochromic properties to either eliminate or facilitate bimodal switching of Chrimson. We will tailor new Chrimson variants with long open states and high proton selectivity for compartmental optogenetics, specifically for depleting protons from synaptic vesicle and lysosome, gaining optogenetic control of their acidification and functionalization.

该项目的目的是进一步表征和设计具有最红移吸收光谱和高质子选择性的通道紫红质，它与古细菌光驱动质子泵和大多数其他通道紫红质具有相同的结构特征。我们将运用生物化学、生物物理和理论等方法对活性位点、外门和选择性过滤器等特殊亚结构进行研究。为了深入了解，我们将建立和研究计算模型，这将有助于解释实验结果并协助合理设计红移变体。我们将设计新的甚至更多的红移变体，增加钠选择性和/或修改动力学。由于在波长超过600纳米时神经元组织的传输增加，这些克里姆森衍生物对神经科学的体内应用最感兴趣。此外，我们将探索生物变色性质，以消除或促进双峰开关。我们将为室室光遗传学量身定制具有长开放状态和高质子选择性的新crimson变体，特别是用于从突触囊泡和溶酶体中消耗质子，获得光遗传学控制其酸化和功能化。