Duchenne muscular dystrophy (DMD) affects 1:3500-5000 newborn boys and manifests with progressive skeletal muscle wasting, respiratory failure and eventual heart failure. Symptoms show different onset from patients' childhood to the second decade of age. We reprogrammed fibroblasts from two independent DMD patients with a complete loss of dystrophin expression, carrying deletions of exons 45-50 and 48-50. The resulting hiPSCs show expression of pluripotency markers (NANOG, OCT4, SSEA4), differentiation capacity into all three germ layers, normal karyotype, genetic identity to the originating parental fibroblasts and the patient-specific dystrophin mutation.

• MeSH
• buněčná diferenciace MeSH
• buněčné linie cytologie   metabolismus   MeSH
• dítě MeSH
• Duchennova muskulární dystrofie genetika   metabolismus   patofyziologie   MeSH
• dystrofin genetika   metabolismus   MeSH
• exony MeSH
• indukované pluripotentní kmenové buňky cytologie   metabolismus   MeSH
• lidé MeSH
• mladiství MeSH
• oktamerní transkripční faktor 3 genetika   metabolismus   MeSH
• sekvenční delece MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Duchenne and Becker muscular dystrophies (DMD/BMD) are associated with mutations in the DMD gene. We determined the mutation status of 47 patients with dystrophinopathy without deletion or duplication in the DMD gene by screening performed by reverse transcription-PCR, protein truncation test, and DNA sequencing. We describe three patients with a mutation creating a premature termination codon (p.E55X, p.E1110X, and p.S3497PfsX2) but with a mild phenotype, which present three different ways of rescuing the DMD phenotype. In one patient we detected the insertion of a repetitive sequence AluYa5 in intron 56, which led to skipping of exon 57. Further, using quantitative analysis of DMD mRNA carrying various mutated alleles, we examine levels of mRNA degradation due to nonsense mediated mRNA decay. The quantity of dystrophin mRNA is different depending on the presence of a mutation leading to a premature termination codon, and position of the analysed mRNA region with respect to its 5' end or 3' end. Average relative amounts of DMD mRNAs carrying a premature termination codon is 48% and 17%, when using primers amplifying the 5' and 3' cDNA regions, respectively.

• MeSH
• bodová mutace MeSH
• Duchennova muskulární dystrofie genetika   MeSH
• dystrofin genetika   metabolismus   MeSH
• exony MeSH
• fenotyp MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• mutační analýza DNA metody   MeSH
• sekvence nukleotidů MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• delece genu MeSH
• dilatační kardiomyopatie genetika   patologie   MeSH
• dystrofin genetika   metabolismus   MeSH
• imunoelektronová mikroskopie metody   MeSH
• křečci praví MeSH
• modely nemocí na zvířatech MeSH
• proteiny genetika   MeSH
• sarkolema metabolismus   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• zvířata MeSH