title: Spatial distribution of transgenic protein after gene electrotransfer to porcine muscle.
abstract: Gene electrotransfer is an effective nonviral technique for delivery of plasmid DNA into tissues. From a clinical perspective, muscle is an attractive target tissue as long-term, high-level transgenic expression can be achieved. Spatial distribution of the transgenic protein following gene electrotransfer to muscle in a large animal model has not yet been investigated. In this study, 17 different doses of plasmid DNA (1-1500 μg firefly luciferase pCMV-Luc) were delivered in vivo to porcine gluteal muscle using electroporation. Forty-eight hours post treatment several biopsies were obtained from each transfection site in order to examine the spatial distribution of the transgenic product. We found a significantly higher luciferase activity in biopsies from the center of the transfection site compared to biopsies taken adjacent to the center, 1 and 2 cm along muscle fiber orientation (p<0.05 and p<0.0001, respectively). On average, 43% of the total luciferase activity was localized in the center biopsy. In conclusion, we found that gene electrotransfer to muscle in a large animal model led to localized gene expression corresponding to the area delineated by the electrodes. High doses of plasmid DNA did not lead to a larger area of the muscle expressing the transgenic protein.
authors.affiliation: Center for Experimental Drug and Gene Electrotransfer, Department of Oncology, Copenhagen University Hospital Herlev, 2730 Herlev, Denmark.
authors.forename: Iben
authors.initials: I
authors.lastname: Spanggaard
authors.forename: Thomas
authors.initials: T
authors.lastname: Corydon
authors.forename: Pernille
authors.initials: P
authors.lastname: Hojman
authors.forename: Hanne
authors.initials: H
authors.lastname: Gissel
authors.forename: Frederik
authors.initials: F
authors.lastname: Dagnaes-Hansen
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Jensen
authors.forename: Julie
authors.initials: J
authors.lastname: Gehl
mesh_terms: D000818:Animals; D018274:Electroporation; D049409:Luciferases, Firefly / Q000235:genetics; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D013552:Swine / Q000378:metabolism
chemical_list: D049409:Luciferases, Firefly