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Use of the Tetracycline System for Inducible Protein Synthesis in the Kidney

Anna Rachel Gallagher^*, Kai Schönig, Nelson Brown^*, Hermann Bujard and Ralph Witzgall

*Institute for Anatomy and Cell Biology I and Center for Molecular Biology, University of Heidelberg, and Institute for Molecular and Cellular Anatomy, University of Regensburg, Germany.

Correspondence to Ralph Witzgall, Institute for Molecular and Cellular Anatomy, University of Regensburg, Universitätsstr. 31, 93053 Regensburg, Germany. Phone: +49-941-9432821; Fax: +49-941-9432868;

ABSTRACT. The great advantage of the tetracycline-inducible system lies in its ability to address a large variety of biological questions in a time-dependent and tissue-specific manner. This study describes a transgenic mouse line, rTA^LAP-1, which produces the reverse tetracycline transactivator under control of the liver activator protein (LAP) promoter. Two reporter lines with luciferase and LacZ reporter genes were used to demonstrate predominant expression in the kidney and liver when doxycycline was added to the drinking water. In the kidney, transgene expression was found primarily in cortical proximal tubules. No luciferase and -galactosidase activity was detected in mice without doxycycline in the drinking water, which attests to the tight control of this system. One of the advantages of the tet system lies in its reversibility, and indeed, a virtually complete remission of transgene activity in both the kidney and liver was observed when doxycycline was withdrawn. Also examined was transactivator activity during development by exposing the mothers producing the reverse transactivator to doxycycline before mating. Transgene activity was detected in newborn kidneys and liver, indicating that sufficient amounts of doxycycline had crossed the placental barrier. During nephron development, the LAP promoter appeared to be only active in the more mature proximal tubules. Finally, the rTA^LAP-1 line was used to inducibly express the human PKD2 cDNA in proximal tubules of transgenic mice, but no cystic changes were detected, even after 6 mo of induction. E-mail: ralph.witzgall@vkl.uni-regensburg.de

Copyright © 2008 by the American Society of Nephrology. Online ISSN: 1533-3450 Print ISSN: 1046-6673