title: Targeted and stable gene delivery into muscle cells by a two-step transfer system.
abstract: We developed a muscle-specific gene delivery system based on two-step gene transfer. The first step involved adenovirus-mediated transfer of the ecotropic retrovirus receptor (EcoRec) gene driven by the muscle-specific desmin promoter. Both human primary myoblasts and fibroblasts were efficiently transduced with this adenovirus vector. However, expression of EcoRec was detected only in myoblasts. In the second step, EcoRec-expressing myoblasts could be stably transduced with the ecotropic retroviral vector with the beta-galactosidase gene. Approximately 15% of myoblasts were transduced by this two-step strategy. When the transduced myoblasts were differentiated into myotubes, extensive cell-cell fusion occurred, and the apparent number of beta-galactosidase-positive cells increased to 28%. These results indicate that our two-step gene delivery system could be used for targeted and stable gene transfer into muscle cells.
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Advanced Medical Technology, Tokyo, 113-8602, Japan.
authors.forename: I
authors.initials: I
authors.lastname: Fujii
authors.forename: S
authors.initials: S
authors.lastname: Suzuki
authors.forename: T
authors.initials: T
authors.lastname: Igarashi
authors.forename: M
authors.initials: M
authors.lastname: Matsukura
authors.forename: T
authors.initials: T
authors.lastname: Miike
authors.forename: T
authors.initials: T
authors.lastname: Shimada
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D002454:Cell Differentiation; D002459:Cell Fusion; D002478:Cells, Cultured; D003893:Desmin / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005347:Fibroblasts; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006651:Histocytochemistry; D006801:Humans; D007223:Infant; D008297:Male; D009132:Muscles / Q000166:cytology / Q000378:metabolism*; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics; D011991:Receptors, Virus / Q000235:genetics* / Q000378:metabolism*; D014161:Transduction, Genetic / Q000235:genetics
chemical_list: D003893:Desmin; D011991:Receptors, Virus