G-protein-coupled receptors (GPCRs) remain major drug targets, despite our incomplete understanding of how they signal through 16 non-visual G-protein signal transducers (collectively named the transducerome) to exert their actions. To address this gap, we have developed an open-source suite of 14 optimized bioluminescence resonance energy transfer (BRET) Gαβγ biosensors (named TRUPATH) to interrogate the transducerome with single pathway resolution in cells. Generated through exhaustive protein engineering and empirical testing, the TRUPATH suite of Gαβγ biosensors includes the first Gα15 and GαGustducin probes. In head-to-head studies, TRUPATH biosensors outperformed first-generation sensors at multiple GPCRs and in different cell lines. Benchmarking studies with TRUPATH biosensors recapitulated previously documented signaling bias and revealed new coupling preferences for prototypic and understudied GPCRs with potential in vivo relevance. To enable a greater understanding of GPCR molecular pharmacology by the scientific community, we have made TRUPATH biosensors easily accessible as a kit through Addgene.

中文翻译：

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TRUPATH，一种用于询问GPCR换能器组的开源生物传感器平台。

尽管我们对G蛋白偶联受体（GPCR）仍是通过16种非可视G蛋白信号转导子（统称为换能器）发挥作用的信号尚不完全了解，但它们仍然是主要的药物靶标。为了解决这个空白，我们开发了一个开放源代码套件，其中包含14种优化的生物发光共振能量转移（BRET）Gαβγ生物传感器（称为TRUPATH），以单细胞分辨率在细胞内询问转导组。通过详尽的蛋白质工程和经验测试生成的TRUPATHGαβγ生物传感器套件包括首批Gα15和GαGustducin探针。在面对面的研究中，在多个GPCR和不同细胞系中，TRUPATH生物传感器的性能优于第一代传感器。使用TRUPATH生物传感器进行的基准研究总结了以前记录的信号偏倚，并揭示了对原型GTP和研究不足的GPCR具有潜在体内相关性的新偶联偏好。为了使科学界对GPCR分子药理学有更深入的了解，我们使TRUPATH生物传感器可以通过Addgene轻松地作为试剂盒获得。