加州大学张瑾小组报道远红色化学遗传激酶生物传感器实现信号网络的多路复用和超分辨率成像。这一研究成果发表在2025年4月21日出版的国际学术期刊《自然—生物技术》上。

在这里，该研究团队介绍了高度敏感的化学遗传激酶活性生物传感器，它结合了遗传可编码的自标记标签HaloTag7和远红色合成荧光团。该技术可实现四维活动成像和功能超分辨率成像，主题是受激排放损耗和其他高分辨率显微镜技术，允许在必要的分辨率下跨尺度检测信号活动。受激发射耗竭成像使研究蛋白激酶A活性在单个网格蛋白包被凹坑。该课题组人员进一步展示了在单个活细胞中多达五种分析物的成像，增加了生物传感器多路复用的维度。细胞对不同G蛋白偶联受体（GPCRs）激活反应的多重成像允许定量测量单个GPCR配体对下游的时空网络状态。

据介绍，荧光生物传感器通过直接测量信号活动的活细胞，促进了生物医学研究。然而，目前的技术提供了有限的分辨率和维度，阻碍了他们解决和询问时空调节的信号活动网络的能力。

附：英文原文

Title: Far-red chemigenetic kinase biosensors enable multiplexed and super-resolved imaging of signaling networks

Author: Frei, Michelle S., Sanchez, Samantha A., He, Xinchang, Liu, Longwei, Schneider, Falk, Wang, Zichen, Hakozaki, Hiroyuki, Li, Yajuan, Lyons, Anne C., Rohm, Theresa V., Olefsky, Jerrold M., Shi, Lingyan, Schneberg, Johannes, Fraser, Scott E., Mehta, Sohum, Wang, Yingxiao, Zhang, Jin

Issue&Volume: 2025-04-21

Abstract: Fluorescent biosensors have advanced biomedical research by enabling direct live-cell measurements of signaling activities. However, current technology offers limited resolution and dimensionality, impeding our ability to resolve and interrogate spatiotemporally regulated networks of signaling activities. Here we introduce highly sensitive chemigenetic kinase activity biosensors that combine the genetically encodable self-labeling tag, HaloTag7, with far-red-emitting synthetic fluorophores. This technology enables both four-dimensional activity imaging and functional super-resolution imaging using stimulated emission depletion and other high-resolution microscopy techniques, permitting signaling activity to be detected across scales with the necessary resolution. Stimulated emission depletion imaging enabled the investigation of protein kinase A activity at individual clathrin-coated pits. We further demonstrate imaging of up to five analytes in single living cells, an increase in the dimensionality of biosensor multiplexing. Multiplexed imaging of cellular responses to the activation of different G-protein-coupled receptors (GPCRs) allowed quantitative measurements of spatiotemporal network states downstream of individual GPCR–ligand pairs.

DOI: 10.1038/s41587-025-02642-8

Source: https://www.nature.com/articles/s41587-025-02642-8