Title | A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression. |
Publication Type | Journal Article |
Year of Publication | 2018 |
Authors | Iwata M, Englund DA, Wen Y, Dungan CM, Murach KA, Vechetti IJ, Mobley CB, Peterson CA, McCarthy JJ |
Journal | Skelet Muscle |
Volume | 8 |
Issue | 1 |
Pagination | 33 |
Date Published | 2018 10 27 |
ISSN | 2044-5040 |
Keywords | Animals, Gene Targeting, Green Fluorescent Proteins, Humans, Mice, Mice, Inbred C57BL, Muscle, Skeletal, PAX7 Transcription Factor, Tetracycline, Trans-Activators, Transgenes |
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. |
DOI | 10.1186/s13395-018-0181-y |
Alternate Journal | Skelet Muscle |
PubMed ID | 30368256 |
PubMed Central ID | PMC6204038 |
Grant List | AR060701 / NH / NIH HHS / United States AR071753 / NH / NIH HHS / United States AG049806 / NH / NIH HHS / United States R01 AR060701 / AR / NIAMS NIH HHS / United States F32 AR071753 / AR / NIAMS NIH HHS / United States R01 AG049806 / AG / NIA NIH HHS / United States TL1 TR001997 / TR / NCATS NIH HHS / United States |