title: shRNA-mediated gene knockdown in skeletal muscle.
abstract: RNA interference appears as a promising tool for therapeutic gene silencing to block protein expression. A long-lived silencing is obtained through the in situ expression of shRNA. A safe approach is to use a physical method such as in vivo electropulsation with plate electrodes. This is presently validated in muscles by the in vivo coelectrotransfer of plasmids specifically coding for expression and silencing of a fluorescent protein. No long-lived tissue damage is observed by the proper choice of the electric pulsing parameters and the amount of injected plasmids. Using a noninvasive fluorescence imaging assay, electrodelivery in mouse muscles is observed to induce complete silencing over more than 2 months in a specific way. The proper choices of the plasmids (sequence, promoter, and relative amounts) appear as key parameters in the successful long-term silencing.
authors: CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), Université de Toulouse, Toulouse, France. muriel.golzio@ipbs.fr
authors: Muriel
authors: M
authors: Golzio
authors: Jean-Michel
authors: JM
authors: Escoffre
authors: Justin
authors: J
authors: Teissié
mesh_terms: D000818:Animals; D018274:Electroporation / Q000379:methods; D005260:Female; D055785:Gene Knockdown Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018482:Muscle, Skeletal / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism*; D059008:Time-Lapse Imaging / Q000379:methods
chemical_list: D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins