In vivo gene electroporation in skeletal muscle with special reference to the duration of gene expression

To determine the limits of the duration of in vivo transferred foreign gene expression, we conducted electroporation (EP), a powerful non-viral means of gene transfer for living animals, into skeletal muscle of rats and mice with a luciferase, GFP or erythropoietin (EPO)-encoding reporter plasmid. The luciferase reporter plasmid was used for optimization of EP conditions, while GFP and EPO plasmids were used for monitoring the duration of gene expression. In the rat, increased hematocrit levels were maintained for at least 9 weeks with approximately a 3-fold increase in plasma EPO protein concentration at 4 weeks post-transfection. In the mouse, the GFP plasmid transfer confirmed that the reporter gene expression lasted as long as 3 months post-transfection. By introducing the EPO gene in vivo in the mouse, increased hematocrit levels revealed that duration of reporter gene expression was at least 14.5 months after in vivo gene EP into skeletal muscle. These results implicate an excellent potential of in vivo gene EP, applicable to both experimental and therapeutic purposes.