Optogenetic Retinal Gene Therapy with the Light Gated GPCR Vertebrate Rhodopsin
Optogenetic Retinal Gene Therapy with the Light Gated GPCR Vertebrate Rhodopsin
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DOI：
10.1007/978-1-4939-7522-8_12
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Flannery, John G.
中科院分区：
文献类型：
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作者：
Gaub, Benjamin M.;Berry, Michael H.;Flannery, John G.
在视网膜疾病中，尽管随着光感受器的死亡而丧失光敏感性，但许多视网膜中间神经元在生理和代谢功能状态下长时间存活。这为从基因上为这些细胞增加光敏功能的治疗提供了机会。光遗传疗法正在开发中，但到目前为止，它们一直存在光敏感度低和微生物视蛋白的动态响应范围窄的问题。几个小组表明，表达光敏G蛋白偶联受体(GPCRs)，如脊椎动物视紫红质，可以通过信号放大增加敏感性。在这里，我们描述了(1)在视网膜神经元中表达光门GPCRs的方法，(2)在体外记录视网膜外植体的光反应，(3)在体内记录皮质光反应，以及(4)在处理的小鼠中测试视觉引导行为。
In retinal disease, despite the loss of light sensitivity as photoreceptors die, many retinal interneurons survive in a physiologically and metabolically functional state for long periods. This provides an opportunity for treatment by genetically adding a light sensitive function to these cells. Optogenetic therapies are in development, but, to date, they have suffered from low light sensitivity and narrow dynamic response range of microbial opsins. Expression of light-sensitive G protein coupled receptors (GPCRs), such as vertebrate rhodopsin, can increase sensitivity by signal amplification, as shown by several groups. Here, we describe the methods to (1) express light gated GPCRs in retinal neurons, (2) record light responses in retinal explants in vitro, (3) record cortical light responses in vivo, and (4) test visually guided behavior in treated mice.