Synergistic effect of ultrasound and PEI on DNA transfection in vitro

Abstract

Ultrasound and poly(ethylenimine) (PEI) have each separately been shown to increase DNA transfection efficiency. This study tested the hypothesis that the combination of ultrasound and PEI can have a synergistic effect to increase DNA transfection. This in vitro study assessed transfection efficiency of two different DNA plasmids encoding green fluorescent protein and firefly luciferase in two different cells types, a primary culture of human aortic smooth muscle cells and an immortal line of human prostrate cancer cells. We found that ultrasound sonication increased transfection up to 18-fold, DNA complexation with PEI increased transfection up to 90-fold, and the combination of ultrasound and PEI synergistically increased transfection up to 200-fold, which resulted in reporter gene expression by 34% of cells. Kinetic measurements found that the effects of ultrasound alone acted quickly, whereas increased transfection by PEI either alone or in combination with ultrasound strongly benefited from a 4-h incubation with the DNA plasmid after sonication. Although serum reduced absolute expression levels, it did not affect the relative increase in transfection when ultrasound was added to PEI enhancement. Flow cytometry measurements showed that sonication increased intracellular uptake of labelled DNA complexed to PEI by 55% relative to PEI complexation alone. Electrophoresis assay showed no damage to DNA or PEI-DNA complexes after sonication. Overall, these results suggest that the combination of ultrasound and PEI can have a synergistic effect to increase DNA transfection.

Efficient gene delivery to myocardium with ultrasound targeted microbubble destruction and polyethylenimine

“…Recent studies have shown that ultrasound-targeted microbubble destruction (UTMD) is a new, non-viral, promising method for the gene delivery to the heart [1][2][3][4][5] . The combination of ultrasound (US) irradiation with polyethylenimine (PEI) can not only improve the transfection efficiency markedly [6,7] , but also achieve specific targeting [8] . Researches showed that UTMD combined with PEI could enhance the transfecZhiyi CHEN, male, born in 1975, Doctorial Candidate E-mai: winchen@smail.hust.edu.cn # Corresponding author * The project was supported by a grant from the National Natural Sciences Foundation of China (No.…”

Ultrasound microbubble contrast agents: Application to therapy for peripheral vascular disease

“…Transfection efficacy in vivo is much lower than that in vitro, 72 although it is increased with exposure of the gene-carrying microbubble to ultrasonic radiation. 43 …”

Localized, non-viral delivery of nucleic acids: Opportunities, challenges and current strategies

“…However, besides use as a contrast agent, these microbubbles have also been recognized as drug delivery vehicles for non-permeable molecules such as macromolecular drugs and NA [114,115]. Deshpande and Prausnitz showed in vitro that the combination of sonoporation using Optison microbubbles and PEI/DNA complexes significantly improved transfection efficiency over each of the treatments alone [116]. Similarly, improved cytoplasmatic uptake of lipoplexes through cell membrane pores was observed [117].…”