Titre :
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Correction of a deep intronic mutation in calpain3 with antisense oligonucleotides in LGMD2A (poster)
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contenu dans :
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Auteurs :
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4th International Congress of Myology, 4ème colloque international de Myologie (9-13 mai 2011; Lille (France)) ;
Blazquez L ;
Aiastui A ;
Pastoriza N ;
Cano A ;
Avril A ;
Garcia L ;
Lopez de Munain Arregui A
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Type de document :
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Article
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Année de publication :
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2011
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Pages :
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p. 112
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Langues:
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Anglais
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Mots-clés :
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colloque
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Résumé :
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Last years several mutations that can be corrected by the exon-skipping technique have been described in Duchenne dystrophy. In LGMD2A, however, all the mutations described to date do not seem to be good candidates for the RNA reparation technology. Our group has identified a deep intronic mutation in LGMD2A that could be potentially repaired at RNA level using a cellular model alternative to myoblasts. Human fibroblasts from a control and from a LGMD2A patient were transduced with a lentiviral vector encoding the cDNA for MyoD and differentiated to myotubes. Three different antisense oligonucleotides targeted to the mentioned mutation were designed and tested in myotubes. Expression of Calpain-3 and other muscle specific genes were analyzed by RTQ-PCR and Western-Blot. The results indicate that Calpain-3 protein is expressed in fibroblasts derived to myotubes by MyoD overexpression in the control but not in the LGMD2A patient. Myotubes of LGMD2A patient treated with antiSD oligonucleotide recover Calpain-3 synthesis at RNA level. Myogenic conversion of Human fibroblasts by MyoD gene transfer could be considered a cellular model alternative to myotubes derived from myoblasts to test ex-vivo gene therapy approaches in calpainopathies, particularly in those patients with a disease progression that makes difficult to obtain a muscle biopsy. Simultaneously, the recovery of Calpain-3 in a LGMD2A patient cell opens a new pathway to treat this disease.
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