Comparative analysis of the effects of ultrasound-targeted microbubble destruction on recombinant adeno-associated virus- and plasmid-mediated transgene expression in human retinal pigment epithelium cells

Abstract

Ultrasound-targeted microbubble destruction (UTMD) has been utilized to deliver a drug/gene into cells in both in vitro and in vivo studies. This study was performed to investigate the feasibility of UTMD-enhanced recombinant adeno-associated virus- (rAAV) and plasmid-mediated transfection into the human retinal pigment epithelium (RPE) cell line ARPE-19. Additionally, the transfection efficiency of rAAV and plasmid was compared in order to choose the appropriate gene vector or strategy to be used with UTMD for RPE-based gene modification. rAAV or plasmid encoding an enhanced green fluorescent protein gene was administered to ARPE-19 cells under various UTMD conditions. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl-tetrazolium bromide (MTT) assay. Transfection efficiency was determined by fluorescence microscopy and flow cytometry. UTMD significantly enhanced the transfection efficiency of rAAV and plasmid in ARPE-19 cells without adverse effects on cell viability. The transfection efficiency of rAAV was higher than that of plasmid. rAAV is therefore an appropriate gene vector for use with UTMD in retinal gene modification.