A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression

Masahiro Iwata, Davis A. Englund, Yuan Wen, Cory M. Dungan, Kevin A. Murach, Ivan J. Vechetti, Christopher B. Mobley, Charlotte A. Peterson, John J. McCarthy

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

Background: The tetracycline-responsive system (Tet-ON/OFF) has proven to be a valuable tool for manipulating gene expression in an inducible, temporal, and tissue-specific manner. The purpose of this study was to create and characterize a new transgenic mouse strain utilizing the human skeletal muscle α-actin (HSA) promoter to drive skeletal muscle-specific expression of the reverse tetracycline transactivator (rtTA) gene which we have designated as the HSA-rtTA mouse. Methods: To confirm the HSA-rtTA mouse was capable of driving skeletal muscle-specific expression, we crossed the HSA-rtTA mouse with the tetracycline-responsive histone H2B-green fluorescent protein (H2B-GFP) transgenic mouse in order to label myonuclei. Results: Reverse transcription-PCR confirmed skeletal muscle-specific expression of rtTA mRNA, while single-fiber analysis showed highly effective GFP labeling of myonuclei in both fast- and slow-twitch skeletal muscles. Pax7 immunohistochemistry of skeletal muscle cross-sections revealed no appreciable GFP expression in satellite cells. Conclusions: The HSA-rtTA transgenic mouse allows for robust, specific, and inducible gene expression across muscles of different fiber types. The HSA-rtTA mouse provides a powerful tool to manipulate gene expression in skeletal muscle.

Original languageEnglish
Article number33
JournalSkeletal Muscle
Volume8
Issue number1
DOIs
StatePublished - Oct 27 2018

Bibliographical note

Funding Information:
This work was supported by NIH grants AG049806 and AR060701 to JJM and CAP and AR071753 to KAM.

Publisher Copyright:
© 2018 The Author(s).

Keywords

  • Skeletal muscle-specific
  • Tetracycline-responsive

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine
  • Molecular Biology
  • Cell Biology

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