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Transgenic Research

, Volume 12, Issue 1, pp 33–43 | Cite as

Tetracycline-Inducible System for Regulation of Skeletal Muscle-Specific Gene Expression in Transgenic Mice

  • Mischala A. Grill
  • Mark A. Bales
  • Amber N. Fought
  • Kristopher C. Rosburg
  • Stephanie J. Munger
  • Parker B. Antin
Article

Abstract

Tightly regulated control of over-expression is often necessary to study one aspect or time point of gene function and, in transgenesis, may help to avoid lethal effects and complications caused by ubiquitous over-expression. We have utilized the benefits of an optimized tet-on system and a modified muscle creatine kinase (MCK) promoter to generate a skeletal muscle-specific, doxycycline (Dox) controlled over-expression system in transgenic mice. A DNA construct was generated in which the codon optimized reverse tetracycline transactivator (rtTA) was placed under control of a skeletal muscle-specific version of the mouse MCK promoter. Transgenic mice containing this construct expressed rtTA almost exclusively in skeletal muscles. These mice were crossed to a second transgenic line containing a bi-directional promoter centered on a tet responder element driving both a luciferase reporter gene and a tagged gene of interest; in this case the calpain inhibitor calpastatin. Compound hemizygous mice showed high level, Dox dependent muscle-specific luciferase activity often exceeding 10,000-fold over non-muscle tissues of the same mouse. Western and immunocytochemical analysis demonstrated similar Dox dependent muscle-specific induction of the tagged calpastatin protein. These findings demonstrate the effectiveness and flexibility of the tet-on system to provide a tightly regulated over-expression system in adult skeletal muscle. The MCKrtTA transgenic lines can be combined with other transgenic responder lines for skeletal muscle-specific over-expression of any target gene of interest.

doxycycline muscle creatine kinase skeletal muscle tet-on transgenic mice 

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Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Mischala A. Grill
    • 1
  • Mark A. Bales
    • 1
  • Amber N. Fought
    • 1
  • Kristopher C. Rosburg
    • 1
  • Stephanie J. Munger
    • 1
  • Parker B. Antin
    • 1
  1. 1.Department of Cell Biology and AnatomyTucsonUSA

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