Color-tuned Channelrhodopsins for Multiwavelength Optogenetics
Color-tuned Channelrhodopsins for Multiwavelength Optogenetics
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DOI：
10.1074/jbc.m112.391185
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发表时间：
2012-09-14
影响因子：
4.8
通讯作者：
Hegemann, Peter
中科院分区：
文献类型：
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作者：
Prigge, Matthias;Schneider, Franziska;Hegemann, Peter
通道视紫红质-2是一种光门控离子通道，是光遗传学研究的重要工具。它被用来通过蓝光控制神经元活动。本文以衣藻channel rhodopin -1和涡旋channel rhodopin -1的嵌合体为基础，构建了可调色的高效通道视紫红质（ChRs）。这些变体表现出极好的表达和质膜整合，导致HEK细胞中的光电流比通道视紫红质-2大3倍。进一步的分子工程得到了吸收最大值为526 ~ 545 nm的嵌合变体，与通道紫红质-2衍生物的吸收最大值为461 ~ 492 nm的嵌合变体吻合良好。额外的动力学微调导致衍生物的开放状态的寿命范围从19毫秒到5秒。最后，结合吸收绿色和蓝色的变体，可以在560和405 nm处独立激活两个不同的神经细胞群。这种新的通道视紫红质变体可能作为神经回路中双色细胞刺激的重要工具箱元素。
Channelrhodopsin-2 is a light-gated ion channel and a major tool of optogenetics. It is used to control neuronal activity via blue light. Here we describe the construction of color-tuned high efficiency channelrhodopsins (ChRs), based on chimeras of Chlamydomonas channelrhodopsin-1 and Volvox channelrhodopsin-1. These variants show superb expression and plasma membrane integration, resulting in 3-fold larger photocurrents in HEK cells compared with channelrhodopsin-2. Further molecular engineering gave rise to chimeric variants with absorption maxima ranging from 526 to 545 nm, dovetailing well with maxima of channelrhodopsin-2 derivatives ranging from 461 to 492 nm. Additional kinetic fine-tuning led to derivatives in which the lifetimes of the open state range from 19 ms to 5 s. Finally, combining green-with blue-absorbing variants allowed independent activation of two distinct neural cell populations at 560 and 405 nm. This novel panel of channelrhodopsin variants may serve as an important toolkit element for dual-color cell stimulation in neural circuits.