Janicot Remi, Garcia-Marcos Mikel
Department of Biochemistry & Cell Biology, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, USA.
Department of Biochemistry & Cell Biology, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, USA; Department of Biology, College of Arts & Sciences, Boston University, Boston, MA 02115, USA.
STAR Protoc. 2024 Dec 20;5(4):103355. doi: 10.1016/j.xpro.2024.103355. Epub 2024 Oct 1.
ONE vector G protein Optical (ONE-GO) biosensors can measure the activity of endogenously expressed G protein-coupled receptors (GPCRs) in primary cells. By detecting G proteins that belong to all four families (G, G, G, G) across cell types, these biosensors provide high experimental versatility. We first describe steps to express ONE-GO biosensors in primary cells using lentiviral transduction. We then detail how to carry out measurements and subsequent analysis to quantify changes in bioluminescence resonance energy transfer (BRET) reporting on endogenous GPCR activity. For complete details on the use and execution of this protocol, please refer to Janicot et al..
单向量G蛋白光学（ONE-GO）生物传感器可测量原代细胞中内源性表达的G蛋白偶联受体（GPCR）的活性。通过检测跨细胞类型属于所有四个家族（Gαs、Gαi/o、Gαq/11、Gα12/13）的G蛋白，这些生物传感器具有很高的实验通用性。我们首先描述使用慢病毒转导在原代细胞中表达ONE-GO生物传感器的步骤。然后，我们详细说明如何进行测量和后续分析，以量化报告内源性GPCR活性的生物发光共振能量转移（BRET）的变化。有关本方案使用和执行的完整详细信息，请参考雅尼科特等人的文献。
