Detection of genome-edited and endogenously expressed G protein-coupled receptors

Abstract

G protein-coupled receptors (GPCRs) are the largest family of membrane receptors and major targets for FDA-approved drugs. The ability to quantify GPCR expression and ligand binding characteristics in different cell types and tissues is therefore important for drug discovery. The advent of genome editing along with developments in fluorescent ligand design offers exciting new possibilities to probe GPCRs in their native environment. This review provides an overview of the recent technical advances employed to study the localisation and ligand binding characteristics of genome-edited and endogenously expressed GPCRs.

Keywords: CRISPR/Cas9; GPCRs; advanced imaging; endogenous; fluorescent ligand; nanobodies.

Fig. 1

New approaches to tag and study endogenous GPCRs in living cells. These include the use of antibodies, nanobodies, fluorescent ligands and CRISPR/Cas9 genome editing in combination with confocal microscopy, bioluminescence microscopy, fluorescence correlation spectroscopy (FCS), bioluminescence (BRET) and time‐resolved Förster (TR‐FRET) resonance energy transfer, fluorescence‐activated cell sorting (FACS), highly inclined and laminated optical sheet (HILO) microscopy and the application of NanoBiT complementation. Figure prepared in ©BioRender (www.biorender.com).