{"doi":"10.1101/2023.03.02.530839","title":"An accessible and generalizable in vitro luminescence assay for detecting GPCR activation","abstract":"ABSTRACT G-protein coupled receptors (GPCRs) serve critical physiological roles as the most abundant family of receptors. Here we describe the design of a generalizable and accessible I n vitro G PCR split N anoLuc l i gand T riggered R eporter ( IGNiTR ), having broad and diverse applications. IGNiTR leverages the interaction between a conformationspecific binder and agonist-activated GPCR to reconstitute a split nanoluciferase. We have demonstrated IGNiTR with three G s -coupled GPCRs and a G i -coupled GPCR with three classes of conformation-specific binders: nanobodies, miniG proteins, and G-protein peptidomimetics. IGNiTR demonstrated binding efficacy and potency values of various Dopamine Receptor D1 (DRD1) ligands that agree well with reported values. IGNiTR also allows the use of a synthetic G protein peptidomimetic, providing easily standardized reagents for characterizing GPCRs and ligands. We demonstrated three applications of IGNiTR: 1) characterizing GPCR functionality during Nanodisc-based reconstitution process; 2) highthroughput screening of ligands against DRD1; 3) detection of opioids for in the field applications. Due to its convenience, accessibility and consistency, IGNiTR will find extensive applications in GPCR ligand detection, screening and GPCR characterization.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":398626,"datarank":0.10397207708399181,"base_score":0.6931471805599453,"endowment":0.6931471805599453,"self_citation_contribution":0.10397207708399181,"citation_network_contribution":0.0,"self_endowment_contribution":0.10397207708399181,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9594,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1174713,"name":"Jennifer Sescil","orcid":"0000-0001-7424-8197","position":1,"is_corresponding":false},{"id":1174714,"name":"Marina C. Sarcinella","orcid":"0000-0002-8284-2855","position":2,"is_corresponding":false},{"id":341256,"name":"Ryan C. Bailey","orcid":"0000-0003-1021-4267","position":3,"is_corresponding":false},{"id":854222,"name":"Wenjing Wang","orcid":"0000-0001-6025-9848","position":4,"is_corresponding":false},{"id":1174712,"name":"Ruby M. Miller","orcid":"0000-0003-4075-2277","position":0,"is_corresponding":true}],"reference_count":48,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:19:47.744358Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}