title: Development of a cell-based assay for screening of phosphodiesterase 10A (PDE10A) inhibitors using a stable recombinant HEK-293 cell line expressing high levels of PDE10A.
abstract: The cDNA encoding PDE10A (phosphodiesterase 10A) was cloned and a stable recombinant HEK-293 (human embryonic kidney-293) cell line expressing high levels of PDE10A was generated. Transient transfection of pCRE-Luc plasmid, harbouring the luciferase reporter gene under the control of CRE (cAMP-response element)-binding sequence, into the stable recombinant cell line, followed by treatment with PDE10 inhibitor, resulted in a dose-dependent increase in luciferase activity. This method provides a simple and sensitive cell-based assay for screening of PDE10 inhibitors for development of novel therapeutics for the treatment of neurological disorders.
authors.affiliation: Department of Biotechnology, Ranbaxy Research Laboratories, Plot No. 20, Sector 18, Udyog Vihar Industrial Area, Gurgaon-122 001, Haryana, India. roop.singh@ranbaxy.com
authors.forename: Roop Singh
authors.initials: RS
authors.lastname: Bora
authors.forename: Dikshi
authors.initials: D
authors.lastname: Gupta
authors.forename: Renu
authors.initials: R
authors.lastname: Malik
authors.forename: Sonia
authors.initials: S
authors.lastname: Chachra
authors.forename: Pratibha
authors.initials: P
authors.lastname: Sharma
authors.forename: Kulvinder Singh
authors.initials: KS
authors.lastname: Saini
mesh_terms: D001681:Biological Assay / Q000379:methods*; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D006801:Humans; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods; D010726:Phosphodiesterase Inhibitors / Q000008:administration & dosage* / Q000032:analysis; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D010726:Phosphodiesterase Inhibitors; D011994:Recombinant Proteins; C119843:PDE10A protein, human; D010727:Phosphoric Diester Hydrolases