Abstract
G protein-coupled receptors (GPCRs) regulate many aspects of cell biology by serving as membrane-bound sensors that coordinate intracellular processes in response to extracellular stimuli. Equipped with over 800 unique GPCRs, humans utilize complex receptor networks for concerted regulation of biological and physiological processes. However, the contributions of individual GPCRs, their distinct cellular roles, and the biological consequences of receptor-specific sensing are difficult to discern in such complex systems. To overcome this challenge, this work describes the engineering of a cell-based platform uniquely equipped for large-scale GPCR studies while providing intricate details into discrete receptor biology. Combining this platform with CRISPR-based techniques led to several tangential technologies enabling more specialized, sub-molecular studies as well as expansion into incrementally larger multi-component systems. These efforts led to several conceptual insights into GPCR biology, including the discovery of an unknown GPCR mechanism. This work also demonstrates the platform’s robust ability to identify an array of pharmacological modulators, ranging from ions to proteins and agonists to antagonists. Together, these studies provided insights into fundamental GPCR biology, revealed new modes of GPCR modulation, and provided innovative technologies for continual scientific exploration.