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 skel...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-10-01
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| Series: | Skeletal Muscle |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13395-018-0181-y |
| _version_ | 1819046915263692800 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Masahiro Iwata |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-21T10:52:03Z |
| format | Article |
| id | doaj.art-d7c9b283fa8b40d48bc49dd7646f3dfa |
| institution | Directory Open Access Journal |
| issn | 2044-5040 |
| language | English |
| last_indexed | 2024-12-21T10:52:03Z |
| publishDate | 2018-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Skeletal Muscle |
| spelling | doaj.art-d7c9b283fa8b40d48bc49dd7646f3dfa2022-12-21T19:06:37ZengBMCSkeletal Muscle2044-50402018-10-01811810.1186/s13395-018-0181-yA novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expressionMasahiro Iwata0Davis A. Englund1Yuan Wen2Cory M. Dungan3Kevin A. Murach4Ivan J. Vechetti5Christopher B. Mobley6Charlotte A. Peterson7John J. McCarthy8The Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyThe Center for Muscle Biology, University of KentuckyAbstract 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.http://link.springer.com/article/10.1186/s13395-018-0181-ySkeletal muscle-specificTetracycline-responsive |
| spellingShingle | 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 A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression Skeletal Muscle Skeletal muscle-specific Tetracycline-responsive |
| title | A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression |
| title_full | A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression |
| title_fullStr | A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression |
| title_full_unstemmed | A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression |
| title_short | A novel tetracycline-responsive transgenic mouse strain for skeletal muscle-specific gene expression |
| title_sort | novel tetracycline responsive transgenic mouse strain for skeletal muscle specific gene expression |
| topic | Skeletal muscle-specific Tetracycline-responsive |
| url | http://link.springer.com/article/10.1186/s13395-018-0181-y |
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