title: Suppression of nonsense mutations in Rett syndrome by aminoglycoside antibiotics.
abstract: Rett Syndrome (RTT) is caused in more than 60% of cases by nonsense mutations in the MECP2 gene. So far, no curative therapy for RTT has become available. In other genetic disorders, it has been shown that aminoglycosides can cause a read-through of nonsense mutations with an efficiency of up to 20%. The aim of this study was to evaluate if this therapeutic concept is applicable to RTT. HeLa cells were transfected with eukaryotic expression vectors carrying mutant alleles of frequently occurring MECP2 nonsense mutations that were N-terminally fused to a FLAG tag. Transfected cells were incubated 24 h in the presence of gentamicin. The expression of full-length protein was analyzed by Western blotting and immunofluorescent cell staining. In the presence of gentamicin a read-through varying between 10 and 21.8% was found, depending on the nucleotide sequence context of the nonsense mutations. The full-length protein was located correctly in the nucleus. We have shown that aminoglycoside-mediated read-through of nonsense mutations in the MECP2 gene can be achieved in vitro with efficiency comparable with that seen in other disorders.
authors.affiliation: Department of Pediatrics and Pediatric Neurology, Georg August University, Göttingen, D-37075 Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Brendel
authors.forename: Edith
authors.initials: E
authors.lastname: Klahold
authors.forename: Jutta
authors.initials: J
authors.lastname: Gärtner
authors.forename: Peter
authors.initials: P
authors.lastname: Huppke
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015153:Blotting, Western; D041321:Chromosomes, Human, X; D018389:Codon, Nonsense / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique; D005839:Gentamicins / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000096:biosynthesis / Q000235:genetics*; D014176:Protein Biosynthesis / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015518:Rett Syndrome / Q000235:genetics*; D014162:Transfection
chemical_list: D000900:Anti-Bacterial Agents; D018389:Codon, Nonsense; D005839:Gentamicins; C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2; D011993:Recombinant Fusion Proteins