title: A nonsense mutation-created intraexonic splice site is active in the lymphocytes, but not in the skeletal muscle of a DMD patient.
abstract: Production of semi-functional dystrophin mRNA from the dystrophin gene encoding a premature stop codon has been shown to modify the severe phenotype of Duchenne muscular dystrophy (DMD). In this study, we report the tissue-specific production of semi-functional dystrophin mRNA via activation of a nonsense mutation-created intraexonic splice acceptor site. In a DMD patient a novel nonsense mutation was identified in exon 42. In his lymphocytes semi-functional dystrophin mRNA with a 63-nucleotide deletion in exon 42 (dys-63) was found to be produced. In vitro splicing assay using hybrid minigenes disclosed that the mutation-created intraexonic splice acceptor site was activated. In his skeletal muscle cells, however, only the authentically spliced dystrophin mRNA was found. This finding identifies the modulation of the splicing of muscle dystrophin mRNA in cases of DMD as a potential target for therapeutic strategies to generate a milder phenotype for this disease.
authors.affiliation: Department of Pediatrics, Kobe University Graduate School of Medicine, 7-5-1 Kusunokicho, Chuo, Kobe, Japan. matsuo@kobe-u.ac.jp
authors.forename: Van Khanh
authors.initials: VK
authors.lastname: Tran
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takeshima
authors.forename: Zhujun
authors.initials: Z
authors.lastname: Zhang
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Habara
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Haginoya
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nishiyama
authors.forename: Mariko
authors.initials: M
authors.lastname: Yagi
authors.forename: Masafumi
authors.initials: M
authors.lastname: Matsuo
mesh_terms: D001483:Base Sequence; D002675:Child, Preschool; D018389:Codon, Nonsense; D004252:DNA Mutational Analysis; D005091:Exons / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D008297:Male; D008957:Models, Genetic; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism*; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics*; D034622:RNA Interference; D022821:RNA Splice Sites; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018389:Codon, Nonsense; D022821:RNA Splice Sites; D012333:RNA, Messenger