Department of Ophthalmology, Visual and Anatomical Sciences (OVAS), Wayne State University School of Medicine, Detroit, MI, USA.
Methods Mol Biol. 2021;2191:189-199. doi: 10.1007/978-1-0716-0830-2_12.
Channelrhodopsin (ChR)-based optogenetics is one promising approach to restore vision in photoreceptor degenerative diseases such as retinitis pigmentosa (RP) and age-related macular degeneration (AMD). Currently, a large number of ChRs from different alga species as well as engineered variants are available. They vary with their light response properties like peak sensitive wavelength (λ), current amplitude, and kinetics. Therefore, it is important to choose an appropriate ChR for practical applications, such as vision restoration. Here we describe a standard laboratory protocol for characterizing properties of ChRs in in vitro in human embryonic kidney (HEK) cells. Based on such characterization, we also discuss the criteria for selecting optimal ChRs for optogenetic vision restoration.
基于通道视紫红质（ChR）的光遗传学是恢复光感受器退行性疾病（如色素性视网膜炎（RP）和年龄相关性黄斑变性（AMD））的一种很有前途的方法。目前，有大量来自不同藻类物种的 ChR 以及工程变体可用。它们的光响应特性，如峰值敏感波长（λ）、电流幅度和动力学，有所不同。因此，选择合适的 ChR 对于实际应用，如视力恢复，非常重要。本文描述了一种在人胚肾（HEK）细胞中体外测定 ChR 特性的标准实验室方案。基于这种特性分析，我们还讨论了选择用于光遗传学视力恢复的最佳 ChR 的标准。
