July 1, 1991Journal of Cellular Biochemistry
PCR analysis of dystrophin gene mutation and expression
Key result
PCR analysis successfully distinguished murine dystrophin transcripts from muscle or brain promoters and identified DNA sequence variations between wild-type and mdx mice.
Population
mdx mouse models for Duchenne muscular dystrophy (DMD) and wild-type mice
Design
Preclinical
Authors
Discussion
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Implication
Advances mdx model characterization; leaves open translation to clinical DMD applications.
Key Points
- Develop a polymerase chain reaction assay to evaluate low-abundance dystrophin transcripts, differentiate tissue-specific promoter usage, and detect point mutations in murine models.
- Employed polymerase chain reaction (PCR) to amplify and analyze murine dystrophin mRNA transcripts expressed at low levels in tissue.
- Established an assay targeting the 5' region to distinguish transcripts initiated from the muscle promoter versus the brain promoter.
- Amplified overlapping segments from the 5' end of dystrophin cDNA to sequence and identify genetic variations between wild-type and three mdx mouse strains.
- Successfully differentiated between muscle-derived and brain-derived murine dystrophin transcripts using the targeted PCR assay.
- Identified that the original mdx mouse strain carries a sequence mutation distinct from the mutations present in two newer mdx isolates.
- Demonstrated that three independently derived mdx mutant lineages are distinct and available for modeling Duchenne muscular dystrophy.
Structured PICO
P
Population
mdx mouse models for Duchenne muscular dystrophy (DMD) and wild-type mice
E
Exposure
Polymerase chain reaction (PCR) analysis of dystrophin gene expression and mutation
O
Outcome
Identification of DNA sequence variations and distinguishing murine dystrophin transcripts expressed from muscle or brain promoterssurrogate
PCR analysis successfully distinguishes dystrophin transcripts and identifies distinct mutations across different mdx mouse models of Duchenne muscular dystrophy.
Cite This Study
Chamberlain et al. (1991) studied Duchenne muscular dystrophy (DMD). Polymerase chain reaction (PCR) analysis was evaluated on Identification of DNA sequence variations and expression of murine dystrophin transcripts. PCR analysis successfully distinguished murine dystrophin transcripts from muscle or brain promoters and identified DNA sequence variations between wild-type and mdx mice.