Recombinant HEK293 cell line expressing human PDE7A (phosphodiesterase 7A, accession number NM_002603).

N-terminal FLAG-tagged human PDE7A has been stably expressed in a human embryonic kidney (HEK293) cell line. PDE7A expression was confirmed by Western blotting. The regulation of intracellular level of cAMP by PDE7A in PDE7A stably-expressed HEK293 cells was characterized by a cell-based reporter assay using pCRE-luc reporter vector. pCRE-luc contains a luciferase gene that is under the control of the cAMP response element (CRE). When cells transiently transfected with pCRE-luc reporter were activated by forskolin, the level of cAMP was upregulated in parental HEK293 cells, inducing the expression of the luciferase reporter, whereas hPDE7A-HEK293 cells showed reduction in the level of cAMP that resulted in lowered expression of luciferase. Inhibition of PDE7A activity by BRL 50481, a PDE7A inhibitor, restored the cAMP level, resulting in higher luciferase activity.

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