Chapter title |
Determination of gene promoter activity in skeletal muscles in vivo.
|
---|---|
Chapter number | 27 |
Book title |
Myogenesis
|
Published in |
Methods in molecular biology, January 2012
|
DOI | 10.1007/978-1-61779-343-1_27 |
Pubmed ID | |
Book ISBNs |
978-1-61779-342-4, 978-1-61779-343-1
|
Authors |
Sarah M. Senf, Andrew R. Judge, Senf, Sarah M., Judge, Andrew R. |
Abstract |
The use of nonviral (plasmid DNA) gene delivery into skeletal muscle has increased significantly in recent years. The procedure is used to overexpress wild-type proteins, express mutant proteins, or knock down endogenous proteins. These manipulations can identify the role of a specific protein in muscle cell biology and physiology. The same procedure of plasmid DNA gene delivery can be used to introduce a gene promoter reporter construct. Such constructs contain a defined sequence of a gene promoter that regulates the expression of a "reporter." This reporter is easily measured and reflects the in vivo transcriptional activity of the gene promoter sequence under study. The gene promoter can be mutated at known transcription factor-binding sites, truncated to identify specific regions of the gene promoter that are required for transcription, or introduced into skeletal muscle with an expression plasmid for a protein believed to regulate the gene's transcription. Therefore, the use of such gene promoter reporters allows for an in-depth physiological assessment of the gene's transcriptional regulation. |
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