Institute of Pharmacology and Toxicology, University of Zürich, Zurich, Switzerland.
Medical University of Vienna, Center for Brain Research, Department for Neuronal Cell Biology, Vienna, Austria.
Elife. 2024 Aug 15;13:RP96496. doi: 10.7554/eLife.96496.
Class-B1 G-protein-coupled receptors (GPCRs) are an important family of clinically relevant drug targets that remain difficult to investigate via high-throughput screening and in animal models. Here, we engineered PAClight1, a novel genetically encoded sensor based on a class-B1 GPCR (the human PAC1 receptor, hmPAC1R) endowed with high dynamic range (Δ/ = 1100%), excellent ligand selectivity, and rapid activation kinetics ( = 1.15 s). To showcase the utility of this tool for in vitro applications, we thoroughly characterized and compared its expression, brightness and performance between PAClight1-transfected and stably expressing cells. Demonstrating its use in animal models, we show robust expression and fluorescence responses upon exogenous ligand application ex vivo and in vivo in mice, as well as in living zebrafish larvae. Thus, the new GPCR-based sensor can be used for a wide range of applications across the life sciences empowering both basic research and drug development efforts.
B1 类 G 蛋白偶联受体 (GPCRs) 是一类重要的具有临床相关性的药物靶点家族，它们很难通过高通量筛选和动物模型进行研究。在这里，我们设计了 PAClight1，这是一种新型的基于 B1 类 GPCR（人 PAC1 受体，hmPAC1R）的基因编码传感器，具有高动态范围（Δ/ = 1100%）、优异的配体选择性和快速激活动力学（= 1.15s）。为了展示该工具在体外应用中的实用性，我们对 PAClight1 转染和稳定表达细胞之间的表达、亮度和性能进行了彻底的表征和比较。通过在小鼠体内和体外以及活体斑马鱼幼虫中展示其在外源配体应用时的稳健表达和荧光反应，证明了其在动物模型中的应用。因此，这种新型基于 GPCR 的传感器可用于生命科学的广泛应用，为基础研究和药物开发提供支持。
