Generation of a tightly regulated doxycycline-inducible model for studying mouse intestinal biology

✍ Scribed by Sabrina Roth; Patrick Franken; Wendy van Veelen; Lau Blonden; Lalini Raghoebir; Berna Beverloo; Ellen van Drunen; Ernst J. Kuipers; Robbert Rottier; Riccardo Fodde; Ron Smits

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 456 KB
• Volume: 47
• Category: Article
• ISSN: 1526-954X
• DOI: 10.1002/dvg.20446

✦ Synopsis

Abstract

To develop a sensitive and inducible system to study intestinal biology, we generated a transgenic mouse model expressing the reverse tetracycline transactivator rtTA2‐M2 under control of the 12.4 kb murine Villin promoter. The newly generated Villin‐rtTA2‐M2 mice were then bred with the previously developed tetO‐HIST1H2BJ/GFP model to assess inducibility and tissue‐specificity. Expression of the histone H2B‐GFP fusion protein was observed exclusively upon doxycycline induction and was uniformly distributed throughout the intestinal epithelium. The Villin‐rtTA2‐M2 was also found to drive transgene expression in the developing mouse intestine. Furthermore, we could detect transgene expression in the proximal tubules of the kidney and in a population of alleged gastric progenitor cells. By administering different concentrations of doxycycline, we show that the Villin‐rtTA2‐M2 system drives transgene expression in a dosage‐dependent fashion. Thus, we have generated a novel doxycycline‐inducible mouse model, providing a valuable tool to study the effect of different gene dosages on intestinal physiology and pathology. genesis 47:7–13, 2009. © 2008 Wiley‐Liss, Inc.