Abstract
(Background) Retrovirus based vectors integrate the transgene into the genome of the target cells, which can sustain long-term expression. However, the transduction efficiency has been known to be low. Ultrasound was reported to increase gene expression using plasmid DNA. However, there are no reports which show ultrasound effect to virus-mediated gene transfer efficiency. (Methods) Retroviral mediated gene transfer system was used to transfect cultured 293T cells, bovine aortic endothelial cells (BAECs), rat aortic smooth muscle cells (RASMCs), and rat skeletal muscle myoblasts (L6 cells) with the LacZ gene combined with ultrasound exposure. Skeletal muscle is thought to be easy and favorable protein producing organ by gene transfer. We also investigated the efficiency of retroviral-mediated gene transfer into regenerating skeletal muscle in vivo. (Results) Below 1.0 watts/cm2 and 5 seconds exposure, ultrasound showed no cytotoxicity to 293T cells transduced by retrovirus, and no toxicity to virus itself. At the same condition, ultrasound resulted in significant increases in retrovirus-mediated gene expression at 6.6-fold, 4.8-fold, 2.3-fold, and 3.2-fold in 293T cells, BAECs, RASMCs, and L6 cells respectively. Ultrasound enhanced in vivo retroviral-mediated LacZ gene transfer by direct injection into hind limb skeletal muscle compared with retrovirus only. (Conclusions) Ultrasound-associated local gene therapy has potential for not only plasmid-DNA-, but also viral-mediated gene transfer in vivo and in vitro.
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