PDE7B-HEK293 human recombinant cell line, 1x10(6) cells

Recombinant HEK293 cell line expressing human PDE7B (phosphodiesterase 7B, accession number NM_018945). Format: Each vial contains 1 X 106 cells in 1 ml of 10% DMSO. Applications: Monitor cAMP and PDE7B signaling pathway activity. Screen activators or inhibitors of PDE7B Introduction: Phosphodiesterases (PDEs) regulate the intracellular levels of cAMP and cGMP by hydrolyzing cAMP and cGMP to their inactive 5' monophosphates. These cyclic nucleotides play an important role as second messengers in diverse physiological functions. PDE7 is a cAMP-specific enzyme and two PDE genes (PDE7A and PDE7B) have been identified. PDE7B is widely expressed in various tissues including brain, heart, liver, skeletal muscle, and pancreas. Inhibition of PDE7 activity by its inhibitors leads to elevated intracellular level of cAMP. Functional validation: Human PDE7B has been stably expressed in a human embryonic kidney (HEK293) cell line. PDE7B expression was confirmed by Western blotting. The function of expressed PDE7B was validated by cAMP detection assay. The regulation of intracellular level of cAMP by PDE7B in PDE7B stably-expressed HEK293cells was characterized by a cell-based reporter assay using pCRE-luc reporter vector. pCRE-luc contains the luciferase gene that is under the control of cAMP response element (CRE). Elevation of intracellular cAMP activates cAMP response element binding protein (CREB) to bind CRE and induces the expression of luciferase. Forskolin is commonly used to raise the intracellular level of cAMP in cell physiology studies. Forskolin resensitizes cell receptors by activating the enzyme adenylyl cyclase and increasing cAMP levels. 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 hPDE7B-HEK293 cells showed reduction in the level of cAMP that resulted in lowered expression of luciferase. These data show the stable expression of PDE7B in HEK293 cells.