小柯机器人

科学家开发出用于光声和超分辨成像的遗传编码光切换分子传感器
2021-11-30 15:49

德国亥姆霍兹慕尼黑中心Andre C. Stiel团队开发出用于光声和超分辨成像的遗传编码光切换分子传感器。2021年11月29日,《自然—生物技术》杂志在线发表了这项成果。

研究人员构建了一个基于GCaMP5G的光切换Ca2+传感器原型,可以用405/488nm的光进行切换,并在结构层面上描述了其分子机制,包括荧光蛋白的核心桶状结构与Ca2+受体分子的相互作用的重要性。研究人员展示了培养细胞中Ca2+浓度的超分辨率成像,以及在控制Ca2+条件下小鼠体内植入肿瘤细胞的光声Ca2+成像。最后,研究人员通过构建基于外周结合蛋白和G-蛋白偶联受体的光切换麦芽糖和多巴胺传感器的例子,展示了这一概念的普遍性。

据介绍,可逆光切换的蛋白质对于许多超分辨率荧光显微镜和光声成像方法是必不可少的。然而,它们还没有被用作测量特定分析物在纳米尺度或活体动物组织中分布的传感器。

附:英文原文

Title: Genetically encoded photo-switchable molecular sensors for optoacoustic and super-resolution imaging

Author: Mishra, Kanuj, Fuenzalida-Werner, Juan Pablo, Pennacchietti, Francesca, Janowski, Robert, Chmyrov, Andriy, Huang, Yuanhui, Zakian, Christian, Klemm, Uwe, Testa, Ilaria, Niessing, Dierk, Ntziachristos, Vasilis, Stiel, Andre C.

Issue&Volume: 2021-11-29

Abstract: Reversibly photo-switchable proteins are essential for many super-resolution fluorescence microscopic and optoacoustic imaging methods. However, they have yet to be used as sensors that measure the distribution of specific analytes at the nanoscale or in the tissues of live animals. Here we constructed the prototype of a photo-switchable Ca2+ sensor based on GCaMP5G that can be switched with 405/488-nm light and describe its molecular mechanisms at the structural level, including the importance of the interaction of the core barrel structure of the fluorescent protein with the Ca2+ receptor moiety. We demonstrate super-resolution imaging of Ca2+ concentration in cultured cells and optoacoustic Ca2+ imaging in implanted tumor cells in mice under controlled Ca2+ conditions. Finally, we show the generalizability of the concept by constructing examples of photo-switching maltose and dopamine sensors based on periplasmatic binding protein and G-protein-coupled receptor-based sensors.

DOI: 10.1038/s41587-021-01100-5

Source: https://www.nature.com/articles/s41587-021-01100-5

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:68.164
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex


本期文章:《自然—生物技术》:Online/在线发表

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