Document Detail


Modification of splicing in the dystrophin gene in cultured Mdx muscle cells by antisense oligoribonucleotides.
MedLine Citation:
PMID:  9618164     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Deletions and point mutations in the gene encoding the cytoskeletal protein dystrophin and its isoforms cause either the severe progressive myopathy Duchenne muscular dystrophy (DMD) or the milder Becker muscular dystrophy (BMD), largely depending on whether the reading frame is lost or maintained respectively. Frameshift mutations tend to result in a lack of dystrophin at the sarcolemma, destabilization of the membrane and degeneration of skeletal muscle. The mdx mouse is a valuable animal model of DMD as it bears a nonsense point mutation in exon 23 of the murine DMD gene leading to an absence of dystrophin expression in the muscle sarcolemma and muscular dystrophy. This report represents a novel approach to correct dystrophin deficiency at the post-transcriptional level by transfection of muscle cells with antisense RNA. Essentially, 2'- O -methyl oligoribonucleotides (2'OMeRNA) were delivered to the nuclei of primary mdx myoblasts in culture. Dystrophin expression was observed in the sarcolemma of transfected mdx myotubes after transfection by an oligonucleotide complementary to the 3' splice site of murine dystrophin intron 22. Direct sequencing of RT-PCR products from these cells revealed precise splicing of exon 22 to exon 30, skipping the mutant exon and creating a novel in-frame dystrophin transcript. As patients with comparable in-frame internal deletions show relatively mild myopathic symptoms, this may in the future offer a therapeutic approach for DMD, as well as for other inherited disorders.
Authors:
M G Dunckley; M Manoharan; P Villiet; I C Eperon; G Dickson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Human molecular genetics     Volume:  7     ISSN:  0964-6906     ISO Abbreviation:  Hum. Mol. Genet.     Publication Date:  1998 Jul 
Date Detail:
Created Date:  1999-01-12     Completed Date:  1999-01-12     Revised Date:  2009-09-29    
Medline Journal Info:
Nlm Unique ID:  9208958     Medline TA:  Hum Mol Genet     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  1083-90     Citation Subset:  IM    
Affiliation:
Division of Biochemistry, Royal Holloway University of London, Egham TW20 0EX, UK. m.dunckley@rpms.ac.uk
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Nucleus / genetics,  metabolism
Cells, Cultured
Dystrophin / biosynthesis,  genetics*
Exons / genetics
Gene Expression / drug effects
Introns / genetics
Mice
Mice, Inbred mdx
Muscle, Skeletal / cytology,  drug effects,  metabolism*
Oligonucleotides, Antisense / pharmacology*
Point Mutation
Polyethyleneimine / pharmacology
RNA Splicing / drug effects*
Sarcolemma / metabolism
Transfection
Grant Support
ID/Acronym/Agency:
//Wellcome Trust
Chemical
Reg. No./Substance:
0/Dystrophin; 0/Oligonucleotides, Antisense; 9002-98-6/Polyethyleneimine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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