title: The CRE luc bioluminescence transgenic mouse model for detecting ligand activation of GPCRs.
abstract: Numerous assays have been developed to investigate the interactions between G-protein-coupled receptors (GPCRs) and their ligands since GPCRs are key therapeutic targets. Reporter-based assays using the cAMP response element (CRE) coupled with bioluminescence from a luciferase reporter have been used extensively in vitro with high-throughput screens (HTS) of large chemical compound libraries. We have generated a transgenic mouse model (CRE luc) with a luciferase reporter under the control of a synthetic promoter that contains several CREs, which supports real-time bioimaging of GPCR ligand activity in whole animals, tissues, or primary cells. In the CRE luc model, GPCR signaling through the cAMP pathway can be detected from the target GPCR that is in a native cellular environment with a full complement of associated receptors and membrane constituents. Multiple independent lines have been produced by random integration of the transgene, resulting in tissue expression profiles covering the major organs. The goal of the CRE luc model is to accelerate the transition from HTS to profiling of GPCR small-molecule leads in preclinical animal disease models, as well as define the mechanism of action of GPCR drugs in three experimental formats: primary cells, tissue homogenates, and whole animals.
authors.affiliation: 1Sanofi NA Research and Development, Boston, MA, USA.
authors.forename: Holly
authors.initials: H
authors.lastname: Dressler
authors.forename: Kyriakos
authors.initials: K
authors.lastname: Economides
authors.forename: Sarah
authors.initials: S
authors.lastname: Favara
authors.forename: Nancy N
authors.initials: NN
authors.lastname: Wu
authors.forename: Zhen
authors.initials: Z
authors.lastname: Pang
authors.forename: H Greg
authors.initials: HG
authors.lastname: Polites
mesh_terms: D000818:Animals; D017362:Cyclic AMP Response Element-Binding Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008024:Ligands; D008163:Luminescent Measurements; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000737:chemistry*; D059467:Transcriptome; D014162:Transfection
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D054852:Small Molecule Libraries
keywords: GPCR; bioluminescence; luciferase; reporter mouse; transgenic