A Study On Duchenne Muscular Dystrophy

1735 WordsApr 20, 20167 Pages
1.0 Abstract Duchenne Muscular Dystrophy is a degenerative X-linked recessive disorder usually resulting in death in the late third decade. Mutation of Dystrophin gene at Xp21 disrupts the mRNA reading frame resulting in absent dystrophin protein in muscle cells. Currently no therapy can counteract the disease effectively. Exon skipping with oligonucleotide administration restores the reading frame of the mRNA to produce truncated but functional dystrophin and requires repeated administration which can cause drug accumulation toxicity. Mesoangioblast Stem Cell therapy has shown safety but limited efficacy due to problems of migration and engraftment in patient skeletal muscles. The approach of Cell Mediated Exon Skipping in this project will aim to draw from the strengths of previous strategies while reducing their individual limitations. A lentiviral vector expressing small nuclear RNA (snRNA), which can cause permanent skipping of exon 51, was delivered to patient fibroblasts and myoblasts. The snRNA from one corrected nucleus was expected to spread to surrounding uncorrected nuclei. Corrected cells were tested against a control of healthy human muscle biopsy tissue, using Immunofluorescence and PCR for Dystrophin. The hypothesis of neighbouring nuclei correction was tested by Quantitative PCR comparing expression of Dystrophin with a reporter gene, Green Fluorescent Protein, in cultures of corrected and uncorrected cells at relative ratio of 1:10 and 1:50.
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