title: Chimeric snRNA molecules carrying antisense sequences against the splice junctions of exon 51 of the dystrophin pre-mRNA induce exon skipping and restoration of a dystrophin synthesis in Delta 48-50 DMD cells.
abstract: Deletions and point mutations in the dystrophin gene cause either the severe progressive myopathy Duchenne muscular dystrophy (DMD) or the milder Becker muscular dystrophy, depending on whether the translational reading frame is lost or maintained. Because internal in-frame deletions in the protein produce only mild myopathic symptoms, it should be possible, by preventing the inclusion of specific mutated exon(s) in the mature dystrophin mRNA, to restore a partially corrected phenotype. Such control has been previously accomplished by the use of synthetic oligonucleotides; nevertheless, a significant drawback to this approach is caused by the fact that oligonucleotides would require periodic administrations. To circumvent this problem, we have produced several constructs able to express in vivo, in a stable fashion, large amounts of chimeric RNAs containing antisense sequences. In this paper we show that antisense molecules against exon 51 splice junctions are able to direct skipping of this exon in the human DMD deletion 48-50 and to rescue dystrophin synthesis. We also show that the highest skipping activity was found when antisense constructs against the 5' and 3' splice sites are coexpressed in the same cell.
authors.affiliation: Institute Pasteur Fondazione Cenci-Bolognetti, Department of Genetics and Molecular Biology, University La Sapienza, P.le Aldo Moro 5, 00185 Rome, Italy.
authors.forename: Fernanda Gabriella
authors.initials: FG
authors.lastname: De Angelis
authors.forename: Olga
authors.initials: O
authors.lastname: Sthandier
authors.forename: Barbara
authors.initials: B
authors.lastname: Berarducci
authors.forename: Silvia
authors.initials: S
authors.lastname: Toso
authors.forename: Giuliana
authors.initials: G
authors.lastname: Galluzzi
authors.forename: Enzo
authors.initials: E
authors.lastname: Ricci
authors.forename: Giulio
authors.initials: G
authors.lastname: Cossu
authors.forename: Irene
authors.initials: I
authors.lastname: Bozzoni
mesh_terms: D000818:Animals; D016189:Dystrophin / Q000096:biosynthesis* / Q000235:genetics*; D005091:Exons; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000378:metabolism / Q000628:therapy*; D009865:Oocytes / Q000378:metabolism; D012313:RNA / Q000235:genetics / Q000494:pharmacology; D012322:RNA Precursors / Q000235:genetics; D012326:RNA Splicing; D016372:RNA, Antisense / Q000235:genetics / Q000494:pharmacology; D012342:RNA, Small Nuclear / Q000235:genetics* / Q000494:pharmacology; D017384:Sequence Deletion; D014161:Transduction, Genetic; D014982:Xenopus laevis
chemical_list: D016189:Dystrophin; D012322:RNA Precursors; D016372:RNA, Antisense; D012342:RNA, Small Nuclear; C039919:RNA, recombinant; D012313:RNA