Improved expression of vascular endothelial growth factor by naked DNA in mouse skeletal muscles: implication for gene therapy of ischemic diseases.

We have constructed an expression vector, pCK, that is able to drive high levels of gene expression in the skeletal muscles of mice. pCK contains not only the full length immediate-early (IE) promoter of human cytomegalovirus but also its entire 5' untranslated region upstream from the start codon of the IE gene. In addition, pCK contains the kanamycin resistance gene, but lacks nucleotide sequences unnecessary for its function as a gene delivery vector, allowing the plasmid size to be 3.7 kb. pCK produced significantly higher levels of vascular endothelial growth factor 165 both in vitro and in vivo than the control vector, the structure of which is similar to naked DNA vectors employed in previous gene therapy trials. pCK would not only significantly increase the therapeutic effects of naked DNA gene therapy but also dramatically cut down the costs for production and treatment.