Reporter Gene Assay Systems for the Investigation of G‐protein‐coupled Receptors

2003 ◽  
pp. 73-94
Author(s):  
Michaela C. Dinger ◽  
Annette G. Beck‐Sickinger
Keyword(s):  
G Protein ◽  
Reporter Gene ◽  
Reporter Gene Assay ◽  
G Protein Coupled Receptors ◽  
Gene Assay ◽  
G Protein Coupled

scholarly journals Development of a Generic Dual-Reporter Gene Assay for Screening G-Protein-Coupled Receptors

2005 ◽  
Vol 10 (5) ◽  
pp. 437-446 ◽  
Author(s):  
Toby C. Kent ◽  
Kevin S. J. Thompson ◽  
Louise H. Naylor
Keyword(s):  
G Protein ◽  
Reporter Gene ◽  
High Throughput Screening ◽  
Reporter Gene Assay ◽  
G Protein Coupled Receptors ◽  
Rapid Expansion ◽  
Compound Libraries ◽  
Dual Reporter ◽  
Gene Assay ◽  
G Protein Coupled

Multiple assay formats have been developed for the pharmacological characterization of G-protein-coupled receptors (GPCRs) and for screening orphan receptors. However, the increased pace of target identification and the rapid expansion of compound libraries present the need to develop novel assay formats capable of screeningmultipleGPCRs simultaneously. To address this need, the authors have developed a generic dual-reporter gene assay that can detect ligand activity at 2 GPCRs within the same assay. Two stableHEK293 cell lineswere generated expressing either a firefly ( Photinus) luciferase gene under the control ofmultiple cAMP-response elements (CREs) or a Renillaluciferase gene under the control ofmultiple 12-Otetradecanoylphorbol-13-acetate (TPA)-responsive elements (TREs). Coseeded reporter cells were used to assess ligandbinding activity at bothGβ s-and Gβ q-coupled receptors. By selectively coexpressing receptors with a chimeric G-protein, agonist activitywas assessed atGβ i/o-coupled receptors in combinationwith eitherGβ s-or Gβ q-coupled receptors. The dual-reporter gene assaywas shown to be capable of simultaneously performing duplexed screens for a variety of agonist and/or antagonist combinations. The data generated from the duplexed reporter assays were pharmacologically relevant, and Zβ factor analysis indicated the suitability of both agonist and antagonist screens for use in high-throughput screening.

A single-vector EYFP reporter gene assay for G protein-coupled receptors

2015 ◽  
Vol 476 ◽  
pp. 40-44 ◽  
Author(s):  
Helle Hald ◽  
Boqian Wu ◽  
Lamine Bouakaz ◽  
Morten Meldal
Keyword(s):  
G Protein ◽  
Reporter Gene ◽  
Reporter Gene Assay ◽  
G Protein Coupled Receptors ◽  
Gene Assay ◽  
G Protein Coupled

Development of an intact cell reporter gene β-lactamase assay for G protein-coupled receptors for high-throughput screening

2003 ◽  
Vol 314 (1) ◽  
pp. 16-29 ◽  
Author(s):  
Priya Kunapuli ◽  
Richard Ransom ◽  
Kathy L Murphy ◽  
Doug Pettibone ◽  
Julie Kerby ◽  
...  
Keyword(s):  
G Protein ◽  
Reporter Gene ◽  
High Throughput ◽  
High Throughput Screening ◽  
Intact Cell ◽  
G Protein Coupled Receptors ◽  
G Protein Coupled
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