Abstract
Phosphodiesterases (PDEs) constitute a superfamily of enzymes that plays an important role in signal transduction by catalysing the hydrolysis of cAMP and cGMP. cDNA encoding PDE7A1 subtype was cloned and a stable recombinant HEK 293 cell line expressing high levels of PDE7A1 was generated. Transient transfection of pCRE-Luc plasmid, harboring luciferase reporter gene into the stable recombinant cell line and subsequent treatment with PDE7 inhibitor, resulted in a dose-dependent increase in luciferase activity. This method provides a simple and sensitive cell-based assay for screening of PDE7 selective inhibitors for the treatment of T cell mediated diseases.
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Acknowledgement
We thank Dr. Sunil Khattar, Department of Biotechnology and Dr. Kedar Padmakar Purnapatre, Department of Infectious Diseases, Ranbaxy Research Laboratories, for scientific and technological comments and advice.
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Renu Malik and Roop Singh Bora contributed equally to this work.
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Malik, R., Bora, R.S., Gupta, D. et al. Cloning, stable expression of human phosphodiesterase 7A and development of an assay for screening of PDE7 selective inhibitors. Appl Microbiol Biotechnol 77, 1167–1173 (2008). https://doi.org/10.1007/s00253-007-1230-3
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DOI: https://doi.org/10.1007/s00253-007-1230-3