小柯机器人

芬兰赫尔辛基大学Vladislav V. Verkhusha团队近期取得重要工作进展，他们开发出利用远红光对工程化受体酪氨酸激酶(RTKs)进行光学控制的一般方法。该项研究成果2022年6月9日在线出版于《自然—方法学》杂志上。

研究人员开发了一种通用的方法来设计用远红光进行光学控制的RTKs。他们将细菌光敏色素DrBphP靶向细胞表面，并允许其光诱导的构象变化通过跨膜螺旋传递到细胞内RTK结构域。这些结构的系统优化产生了光学调节的表皮生长因子受体HER2、TrkA、TrkB、FGFR1、IR1、cKIT和cMet，命名为eDrRTKs。eDrRTKs在数十秒内诱导哺乳动物细胞中的下游信号传导。用远红光激活eDrRTK的能力使荧光探针在更短的光谱范围内工作，从而实现全光学分析。

研究人员在小鼠中验证了eDrTrkB的性能，发现新皮质中的微创刺激通过颅骨远红光穿透可以诱导神经活动、早期即时基因表达和影响睡眠模式。

据介绍，调节受体酪氨酸激酶(RTK)活性是研究健康和疾病细胞信号通路的必要条件。

附：英文原文

Title: A general approach for engineering RTKs optically controlled with far-red light

Author: Leopold, Anna V., Thankachan, Stephen, Yang, Chun, Gerashchenko, Dmitry, Verkhusha, Vladislav V.

Issue&Volume: 2022-06-09

Abstract: Regulation of receptor tyrosine kinase (RTK) activity is necessary for studying cell signaling pathways in health and disease. We developed a generalized approach for engineering RTKs optically controlled with far-red light. We targeted the bacterial phytochrome DrBphP to the cell surface and allowed its light-induced conformational changes to be transmitted across the plasma membrane via transmembrane helices to intracellular RTK domains. Systematic optimization of these constructs has resulted in optically regulated epidermal growth factor receptor, HER2, TrkA, TrkB, FGFR1, IR1, cKIT and cMet, named eDrRTKs. eDrRTKs induced downstream signaling in mammalian cells in tens of seconds. The ability to activate eDrRTKs with far-red light enabled spectral multiplexing with fluorescent probes operating in a shorter spectral range, allowing for all-optical assays. We validated eDrTrkB performance in mice and found that minimally invasive stimulation in the neocortex with penetrating via skull far-red light-induced neural activity, early immediate gene expression and affected sleep patterns. A suite of eight optogenetic receptor tyrosine kinases controlled by conformational changes in bacterial phytochrome induced by far-red light is introduced. These optogenetic tools perform well in cells and in the brain of behaving mice.

DOI: 10.1038/s41592-022-01517-z

Source: https://www.nature.com/articles/s41592-022-01517-z

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex