Titre : | In vivo imaging of gene expression inhibition by electrically mediated siRNA delivery (abstract : congrès international de Myologie, 2005) |
contenu dans : | |
Auteurs : | Congrès international de myologie 2005 (International Congress of Myology 2005; 9-13 mai 2005; Nantes, France) ; Golzio M ; Mazzolini L ; Moller P ; Rols MP ; Escoffre JM ; Teissié J |
Type de document : | Article |
Année de publication : | 2005 |
Pages : | p. 103 |
Langues: | Anglais |
Mots-clés : | ARN interférent ; colloque ; électrodiagnostic ; étude quantitative ; expression génique ; in vivo ; microscopie à fluorescence ; muscle squelettique ; souris |
Résumé : |
Communication n° 148. Fluorescence imaging on live animal is a new efficient and reproducible method for in vivo detection of gene expression with time on the same animal. Linked to a gene of interest, natural fluorescent proteins (GFP, dsRed) used as reporters genes, allows the detection of their expression. GFP expression in muscle is obtained after plasmid injection in the tibialis cranialis of an anesthesied mouse followed by train of electric pulses applied with planes electrodes on the skin. Gene expression is detected by observing mouse leg under a fluorescence stereomicroscope. Detection is possible using a digitized camera cooled CCD with short exposing time. Qualitative and quantitative analyses are then possible on live mice (balb/c, C57Bl6) despite technical and specific problems such as autofluorescence. In the present work, in vivo fluorescence imaging was used to monitor that electrically mediated siRNA transfer can suppress transgene expression in vivo in adult mice muscles. Using electropulsation for siRNA delivery opens the way for a targeted gene silencing on a broad range of tissue. Gene silencing was efficiently obtained in vivo in the muscle of an adult mammal (mouse) with the coelectrodelivery of a GFP coding plasmid and of the relevant chemically synthesized siRNA. The associated gene silencing lasted over two weeks in vivo. Gene silencing was obtained not only on young adult mice but also on much older animals. Clinical applications of electropulsation for delivery of other classes of molecules are under trials. No tissue damages are detected under our electrical conditions. Therefore this biophysical method should provide an efficient approach of a localized delivery of siRNAs in muscles. Supports : IPA, ARC, Region Midi-Pyrenees, La ligue contre le cancer. Golzio M, Mazzolini L, Moller P, Rols MP, and Teissie J. Inhibition of gene expression in mice muscle by in vivo electrically mediated siRNA delivery. Gene Ther. 2004 : 1-6. |