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Applications: Useful to monitor cAMP and PDE7B signaling pathway activity and to screen activators or inhibitors of PDE7B.
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 PDE7B (phosphodiesterase 7B, accession number NM_018945).
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. 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.
Format: aqueous solution containing DMSO
Genbank: NM_018945
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: Q9NP56
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: 1. Malik, R. et al. (2008) Cloning, stable expression of human phosphodiesterase 7A and development of an assay for screening of PDE7 selective inhibitors. Appl. Microbiol. Biotechnol. 77 (5): 1167-1173
2. Fan Chung, K. (2006) Phosphodiesterase inhibitors in airways disease. Eur. J. Pharmacol. 533(1-3):110-117
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