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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
Author(s) -
Bora Roop Singh,
Gupta Dikshi,
Malik Renu,
Chachra Sonia,
Sharma Pratibha,
Saini Kulvinder Singh
Publication year - 2008
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20070090
Subject(s) - recombinant dna , phosphodiesterase , hek 293 cells , luciferase , cell culture , complementary dna , microbiology and biotechnology , pde10a , transfection , plasmid , reporter gene , biology , chemistry , gene , biochemistry , gene expression , enzyme , genetics
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.