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: Emmanuel Richard, Gaelle Douillard-Guilloux, Catherine Caillaud
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 709 Pg. 223-35 ( 2011) ISSN: 1940-6029 [Electronic] United States
• PMID: 21194031 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• G protein, vesicular stomatitis virus
• Membrane Glycoproteins
• RNA, Small Interfering
• Viral Envelope Proteins
• Glycogen
• Glycogen Synthase

Topics

• Animals
• Cells, Cultured
• Gene Expression
• Genetic Vectors
• Glycogen (biosynthesis)
• Glycogen Storage Disease Type II (genetics, therapy)
• Glycogen Synthase (genetics)
• Lentivirus (genetics)
• Membrane Glycoproteins (genetics)
• Mice
• Muscular Diseases (therapy)
• Myoblasts
• RNA Interference
• RNA, Small Interfering (genetics, therapeutic use)
• Viral Envelope Proteins (genetics)