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A Novel Tetracycline-Responsive Transgenic Mouse Strain for Skeletal Muscle-Specific Gene Expression
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.
Document Type
Article
Publication Date
10-27-2018
Digital Object Identifier (DOI)
https://doi.org/10.1186/s13395-018-0181-y
Funding Information
This work was supported by NIH grants AG049806 and AR060701 to JJM and CAP and AR071753 to KAM.
Related Content
The datasets used and/or analyzed during the current study are available from the corresponding author on request. The HSA-rtTA mouse is available upon request.
Repository Citation
Iwata, Masahiro; Englund, Davis A.; Wen, Yuan; Dungan, Cory M.; Murach, Kevin A.; Vechetti, Ivan J. Jr.; Mobley, Christopher B.; Peterson, Charlotte A.; and McCarthy, John J., "A Novel Tetracycline-Responsive Transgenic Mouse Strain for Skeletal Muscle-Specific Gene Expression" (2018). Center for Muscle Biology Faculty Publications. 7.
https://uknowledge.uky.edu/musclebiology_facpub/7
Notes/Citation Information
Published in Skeletal Muscle, v. 8, 33, p. 1-8.
© The Author(s). 2018
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