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Applications: Useful to monitor cAMP and PDE7A signaling pathway activity and to screen activators or inhibitors of PDE7A.
Background: Phosphodiesterases (PDEs) regulate the intracellular levels of cAMP and cGMP byhydrolyzing cAMP and cGMP to their inactive 5' monophosphates. These cyclicnucleotides play an important role as second messengers in diverse physiologicalfunctions. PDE7 is a cAMP-specific enzyme and two PDE genes (PDE7A and PDE7B)have been identified. PDE7 is widely expressed in various tissues, with PDE7A foundprimarily in skeletal muscle, T lymphocytes, and pancreas, while high levels of PDE7Bare detected in brain, heart, and liver. Inhibition of PDE7 activity by its inhibitors leads toelevated intracellular level of cAMP.
Description: 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.
Format: aqueous solution containing DMSO
Genbank: NM_002603
Host Cell Line: HEK293
Mycoplasma Testing: The cell line has been screened using the PCR-based Venor™GeM Mycoplasma Detection kit (Sigma Aldrich) to confirm the absence of Mycoplasma species.
Storage Stability: Store in liquid nitrogen immediately upon receipt.
Supplied As: Each vial contains ~1 X 10^6 cells in 1 ml of 10% DMSO.
Uniprot: Q13946
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: 1. Malik, R. et al. (2008) Appl. Microbiol. Biotechnol. 77 (5): 1167-1173. 2. Fan Chung, K. (2006). Eur. J. Pharmacol. 533(1-3):110-117.
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