Abstract
BackgroundThe 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.MethodsTo 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.ResultsReverse 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.ConclusionsThe 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.
Highlights
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
Englund contributed to this work. 1The Center for Muscle Biology, University of Kentucky, Lexington, KY 40536, USA 3Department of Physiology, College of Medicine, University of Kentucky, 800 Rose Street, Medical Science Building, Rm: MS-607A, Lexington, KY 40536, USA Full list of author information is available at the end of the article we generated a transgenic mouse which uses the human skeletal muscle α-actin (HSA) promoter to drive skeletal muscle-specific expression of the reverse-tetracycline transactivator which we have designated as the HSA-reverse tetracycline transactivator (rtTA) mouse
Skeletal muscle-specific expression of rtTA mRNA We determined by reverse transcription-PCR the expression of rtTA mRNA in several hind limb muscles, the diaphragm, fat, and several other non-muscle organs
Summary
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 tetracycline-responsive system (Tet-ON/OFF) has proven to be a powerful tool in biomedical research because of the ability to manipulate gene expression within the mouse in both a temporal and tissue-specific manner [1, 2]. A number of skeletal muscle-specific Tet-ON/OFF mice have been described, they have used promoters that drive primarily fast-twitch, type II gene expression; in addition, these mice are not readily available [3, 4].
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