科研之友
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摘要
Inefficient gene delivery poses a challenge for non-viral gene therapy. Cytoplasmic and nuclear membrane barriers are responsible for the inefficiency as they restrict the import of exogenous genes. The present study aimed to improve the transfection efficiency using a novel gene delivery system, which consisted of two components: ultrasound microbubbles and the nuclear factor kappa B (NF kappa B) binding motif. Ultrasound-targeted microbubble destruction (UTMD) was used to enhance the cytoplasmic import of plasmids and the NF kappa B binding motif was added to promote the nuclear intake of the plasmid from the cytoplasm. In the present study, human umbilical vein endothelial cells were transfected using UTMD with two different Cy3-labeled plasmids, phSDF-1 alpha and phSDF-1 alpha-NF kappa B. phSDF-1 alpha-NF kappa B was constructed by inserting a specific DNA targeting sequence (five optimal repeats of the binding motif for the inducible transcription factor NF kappa B) into phSDF-1 alpha. The nuclear import and gene expression efficiency of phSDF-1 alpha-NF kappa B were compared with those of phSDF-1 alpha to investigate the effect of the NF kappa B binding motif on transfection. The results showed that UTMD significantly increased the cytoplasmic intake of pDNA and maintained high cell viability. The nuclear import and gene expression of phSDF-1 alpha-NF kappa B-transfected cells were significantly higher than those transfected with phSDF-1 alpha. Compared with the NF kappa B-free plasmids, the quantity of NF kappa B plasmids in the nucleus increased 6.5-fold and the expression of SDF-1 alpha was 4.4-fold greater. These results suggest that UTMD combined with NF kappa B binding motif significantly improve transfection efficiency by enhancing the cytoplasmic and nuclear import of exogenous plasmid DNA.
- 出版日期2013-11
- 单位武汉大学
