Wang Chao-Qun, Wu Jin-Long, Zhuo Ren-Xi, Cheng Si-Xue
Key Laboratory of Biomedical Polymers of Ministry of Education, Department of Chemistry, Wuhan University, Wuhan 430072, P. R. China.
Mol Biosyst. 2014 Mar 4;10(3):672-8. doi: 10.1039/c3mb70502a. Epub 2014 Jan 20.
Ternary nanoparticles, protamine sulfate-calcium carbonate-plasmid DNA (PS-CaCO3-DNA), were prepared for efficient gene delivery. By adding the cationic polypeptide PS in the co-precipitation system of calcium carbonate and DNA, PS-CaCO3-DNA nanoparticles could be formed by self-assembly facilely. The effect of PS on the properties of the ternary nanoparticles was studied by varying the PS amount in the nanoparticles. The size and ζ-potential measurements indicated that the ternary nanoparticles with an appropriate PS amount exhibited a decreased size and an increased ζ-potential. The in vitro gene transfections mediated by different nanoparticles in 293T cells and HeLa cells were carried out in the presence of 10% fetal bovine serum, using pGL3-Luc and pEGFP-C1 as reporter plasmids. As compared with both PS-DNA nanoparticles and CaCO3-DNA nanoparticles, PS-CaCO3-DNA nanoparticles exhibited significantly enhanced gene delivery efficiency, which was higher than that of Lipofectamine 2000-DNA. Confocal microscopy observation showed that PS-CaCO3-DNA nanoparticles could efficiently deliver DNA to cell nuclei. These results indicated that the ternary PS-CaCO3-DNA nanoparticles prepared in this study have promising applications in gene delivery.
制备了三元纳米颗粒——硫酸鱼精蛋白-碳酸钙-质粒DNA（PS-CaCO3-DNA），用于高效基因递送。通过在碳酸钙与DNA的共沉淀体系中加入阳离子多肽PS，PS-CaCO3-DNA纳米颗粒能够通过自组装轻松形成。通过改变纳米颗粒中PS的量，研究了PS对三元纳米颗粒性质的影响。尺寸和ζ电位测量表明，具有合适PS量的三元纳米颗粒尺寸减小且ζ电位增加。在含有10%胎牛血清的情况下，使用pGL3-Luc和pEGFP-C1作为报告质粒，在293T细胞和HeLa细胞中进行了不同纳米颗粒介导的体外基因转染。与PS-DNA纳米颗粒和CaCO3-DNA纳米颗粒相比，PS-CaCO3-DNA纳米颗粒表现出显著提高的基因递送效率，高于Lipofectamine 2000-DNA。共聚焦显微镜观察表明，PS-CaCO3-DNA纳米颗粒能够有效地将DNA递送至细胞核。这些结果表明，本研究制备 的三元PS-CaCO3-DNA纳米颗粒在基因递送方面具有广阔的应用前景。
