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dc.contributor.authorGrose, William E.
dc.contributor.authorClark, K. Reed
dc.contributor.authorGriffin, Danielle
dc.contributor.authorMalik, Vinod
dc.contributor.authorShontz, Kimberly M.
dc.contributor.authorMontgomery, Chrystal L.
dc.contributor.authorLewis, Sarah
dc.contributor.authorBrown, Robert H.
dc.contributor.authorJanssen, Paul M. L.
dc.contributor.authorMendell, Jerry R.
dc.contributor.authorRodino-Klapac, Louise R.
dc.contributor.editorDu, Quansheng
dc.date.accessioned2012-10-26T16:40:51Z
dc.date.available2012-10-26T16:40:51Z
dc.date.issued2012-06-15en_US
dc.identifier.citationPLoS One. 2012 Jun 15; 7(6):e39233en_US
dc.identifier.issn1932-6203en_US
dc.identifier.pmid22720081en_US
dc.identifier.doi10.1371/journal.pone.0039233en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/714
dc.description.abstractThe dysferlinopathies comprise a group of untreatable muscle disorders including limb girdle muscular dystrophy type 2B, Miyoshi myopathy, distal anterior compartment syndrome, and rigid spine syndrome. As with other forms of muscular dystrophy, adeno-associated virus (AAV) gene transfer is a particularly auspicious treatment strategy, however the size of the DYSF cDNA (6.5 kb) negates packaging into traditional AAV serotypes known to express well in muscle (i.e. rAAV1, 2, 6, 8, 9). Potential advantages of a full cDNA versus a mini-gene include: maintaining structural-functional protein domains, evading protein misfolding, and avoiding novel epitopes that could be immunogenic. AAV5 has demonstrated unique plasticity with regards to packaging capacity and recombination of virions containing homologous regions of cDNA inserts has been implicated in the generation of full-length transcripts. Herein we show for the first time in vivo that homologous recombination following AAV5.DYSF gene transfer leads to the production of full length transcript and protein. Moreover, gene transfer of full-length dysferlin protein in dysferlin deficient mice resulted in expression levels sufficient to correct functional deficits in the diaphragm and importantly in skeletal muscle membrane repair. Intravascular regional gene transfer through the femoral artery produced high levels of transduction and enabled targeting of specific muscle groups affected by the dysferlinopathies setting the stage for potential translation to clinical trials. We provide proof of principle that AAV5 mediated delivery of dysferlin is a highly promising strategy for treatment of dysferlinopathies and has far-reaching implications for the therapeutic delivery of other large genes.
dc.rightsGrose et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_US
dc.subjectResearch Articleen_US
dc.subjectBiologyen_US
dc.subjectGeneticsen_US
dc.subjectHuman Geneticsen_US
dc.subjectGene Therapyen_US
dc.subjectGenomicsen_US
dc.subjectGenomic Medicineen_US
dc.subjectGene Therapyen_US
dc.subjectMolecular Cell Biologyen_US
dc.subjectCellular Typesen_US
dc.subjectMuscle Fibersen_US
dc.subjectMedicineen_US
dc.subjectClinical Geneticsen_US
dc.subjectGene Therapyen_US
dc.subjectClinical Research Designen_US
dc.subjectAnimal Models of Diseaseen_US
dc.subjectPreclinical Modelsen_US
dc.subjectNeurologyen_US
dc.subjectMuscular Dystrophiesen_US
dc.subjectNeuromuscular Diseasesen_US
dc.titleHomologous Recombination Mediates Functional Recovery of Dysferlin Deficiency following AAV5 Gene Transferen_US
dc.typeArticleen_US
dc.identifier.pmcidPMC3376115en_US
dc.contributor.corporatenameDepartment of Neurology
dc.contributor.corporatenameCollege of Graduate Studies
refterms.dateFOA2019-04-10T00:39:32Z
html.description.abstractThe dysferlinopathies comprise a group of untreatable muscle disorders including limb girdle muscular dystrophy type 2B, Miyoshi myopathy, distal anterior compartment syndrome, and rigid spine syndrome. As with other forms of muscular dystrophy, adeno-associated virus (AAV) gene transfer is a particularly auspicious treatment strategy, however the size of the DYSF cDNA (6.5 kb) negates packaging into traditional AAV serotypes known to express well in muscle (i.e. rAAV1, 2, 6, 8, 9). Potential advantages of a full cDNA versus a mini-gene include: maintaining structural-functional protein domains, evading protein misfolding, and avoiding novel epitopes that could be immunogenic. AAV5 has demonstrated unique plasticity with regards to packaging capacity and recombination of virions containing homologous regions of cDNA inserts has been implicated in the generation of full-length transcripts. Herein we show for the first time in vivo that homologous recombination following AAV5.DYSF gene transfer leads to the production of full length transcript and protein. Moreover, gene transfer of full-length dysferlin protein in dysferlin deficient mice resulted in expression levels sufficient to correct functional deficits in the diaphragm and importantly in skeletal muscle membrane repair. Intravascular regional gene transfer through the femoral artery produced high levels of transduction and enabled targeting of specific muscle groups affected by the dysferlinopathies setting the stage for potential translation to clinical trials. We provide proof of principle that AAV5 mediated delivery of dysferlin is a highly promising strategy for treatment of dysferlinopathies and has far-reaching implications for the therapeutic delivery of other large genes.


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