Engineering, applications, and future perspectives of GPCR ‐based genetically encoded fluorescent indicators for neuromodulators

This study provides a comprehensive review of genetically encoded fluorescent indicators (GEFIs) based on G-protein-coupled receptors (GPCRs). It discusses the development, structure, and engineering strategies of GEFIs, emphasizing the importance of both sensing (GPCR structure and activation) and reporting (fluorescent protein structure and modulation) domains. The review highlights the applications in measuring neuromodulators with high specificity and spatiotemporal resolution, addresses current limitations and misconceptions in the field, and suggests future directions for enhancing ligand-binding properties and spectral diversity of GEFIs. AbstractThis review explores the evolving landscape of G-protein-coupled receptor (GPCR)-based genetically encoded fluorescent indicators (GEFIs), with a focus on their development, structural components, engineering strategies, and applications. We highlight the unique features of this indicator class, emphasizing the importance of both the sensing domain (GPCR structure and activation mechanism) and the reporting domain (circularly permuted fluorescent protein (cpFP) structure and fluorescence modulation). Further, we discuss indicator engineering approaches, including the selection of suitable cpFPs and expression systems. Additionally, we showcase the diversity and flexibility of their application by presenting a summary of studies where such indicators were used. Along with all the advantages, we also focus on the current limitat...
Source: Journal of Neurochemistry - Category: Neuroscience Authors: Tags: REVIEW Source Type: research