Visual function restoration with a highly sensitive and fast Channelrhodopsin in blind mice

作者：Fei Chen, Xiaodong Duan, Yu Yao, Shang Fa Yang, Yuanyuan Chen, Christine E. Gee, Georg Nagel, Kang Zhang, Shiqiang Gao, Yin Shen · 发表于：Signal Transduction and Targeted Therapy · 年份：2022 · DOI：10.1038/s41392-022-00935-x · 被引用次数：38 · 研究领域：Photoreceptor and optogenetics research、Neuroscience and Neural Engineering、Neuroscience and Neuropharmacology Research

Inherited and age-related retinal degenerative diseases cause progressive loss of photoreceptors, ultimately leading to blindness. Optogenetics is a promising strategy for restoring visual function through photosensitive proteins’ ectopic expression in surviving retinal neurons. 1 Very recently, the optogenetic method with a red-shifted Channelrhodopsin was clinically applied for partial recovery of visual function in a blind patient. 2 However, major obstacles to achieving optimal optogenetic vision restoration are either the low light sensitivity or the slow kinetics of existing rhodopsin-based optogenetic tools, which can be improved by molecular engineering to enhance the efficacy of fast Channelrhodopsins (ChRs). Here, we present a newly engineered ChR variant Ps CatCh2.0, engineered from Ps ChR, 3 which displays inherently high Ca 2+ and Na + conductance and fast kinetics. 3 , 4 We introduced a novel mutation Ps ChR L115C ( Ps CatCh) to enhance its Ca 2+ and Na + permeability further and fused the cleavable N-terminal signal peptide Lucy-Rho (LR 5 in Fig. 1a ), in addition to a plasma membrane trafficking signal (T) and ER export signal (E), to improve its expression and plasma membrane targeting. Ps CatCh2.0 exhibited significant improvements in expression levels/plasma membrane targeting efficiency and a larger photocurrent (Fig. 1a, b, e ). 100-fold less light intensity is needed to generate a similar photocurrent response with Ps CatCh2.0 than with CatCh (Fig. 1b ),...