title: Lentiviral vector delivery of shRNA into cultured primary myogenic cells: a tool for therapeutic target validation.
abstract: RNA interference has emerged as a powerful technique to down-regulate gene expression. The lentiviral vector-mediated expression of small hairpin RNAs (shRNAs) from polymerase III promoters allows permanent down-regulation of a specific gene in a wide range of cell types both in vitro and in vivo. In this chapter, we describe a method permitting the expression of shRNA from lentiviral vectors in primary murine myogenic cells. We designed shRNAs targeted to the muscular glycogen synthase isoform (shGYS1), a highly regulated enzyme responsible for glycogen synthesis, in order to modulate the muscle glycogen biosynthetic pathway and to improve the phenotype in primary myogenic cells from a murine model of glycogen storage disease type II (GSDII). This method based on shRNA-mediated down-regulation could be applied to other muscular disorders to evaluate new therapeutic options.
authors.affiliation: INSERM U876, IFR 66, Université Bordeaux 2, 46 rue Léo Saignat, Bordeaux, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Richard
authors.forename: Gaelle
authors.initials: G
authors.lastname: Douillard-Guilloux
authors.forename: Catherine
authors.initials: C
authors.lastname: Caillaud
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D006003:Glycogen / Q000096:biosynthesis; D006009:Glycogen Storage Disease Type II / Q000235:genetics / Q000628:therapy; D006006:Glycogen Synthase / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D009135:Muscular Diseases / Q000628:therapy; D032446:Myoblasts; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000627:therapeutic use; D014759:Viral Envelope Proteins / Q000235:genetics
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D034741:RNA, Small Interfering; D014759:Viral Envelope Proteins; D006003:Glycogen; D006006:Glycogen Synthase