摘要
目的:寻找一种转染效率高,细胞毒性低的非病毒基因载体,研究以人体内源性精胺为单体,以乙二醇二氯甲酸酯作为连接剂,以聚乙二醇(PEG)作为亲水基团连接剂合成亲水修饰聚阳离子载体PEG-Polycarbam-SP的基因担载效率,以及对非洲绿猴肾癌细胞COS-7的转染活性和细胞毒性的影响。方法:琼脂糖凝胶电泳方法考察复合物的基因担载效率,检测基因复合物的粒径和电位,以荧光素酶质粒为报告基因,研究PEG-Polycarbam-SP/DNA的复合物在COS-7细胞的转染活性,用MTT方法研究PEG-Polycarbam-SP对COS-7细胞的毒性。结果:聚合物与质粒在质量比5以后形成的复合物粒径稳定在50nm左右,Zate电位在20mV左右。COS-7细胞实验显示PEG-Polycarbam-SP具有低于PEI 25kDa的细胞毒性,同时也具有高效输送DNA的能力。结论:PEG-Polycarbam-SP是一种新型的高效、低毒,在基因治疗领域有潜在应用价值的非病毒基因输送载体。
Objective： To find a non-viral gene vector with high transfection efficiency and low cytotoxicity. And to investigate the gene-bound efficiency, transfection efficiency and cell cytotoxicity of the PEG-modified cationic gene vector. PEG-Polycarbam-SP is synthesized by using human endogenous spermine as monomer and ethylenebis（chloroformate） as coupling agent and PEG as hydrophilic coupling agent. Methods： Gene-bound efficiency was examined by AGE（agarose gel electrophoresis）, then particle size and zeta potential of the complexes were measured. Luciferase plasmid was used as the reporter gene to investigate transfection efficiency on COS-7 cells. Cytotoxicity was detected by MTT method. Results： The AGE showed that the plasmid was fully protected at a ratio of 5：1w/w. The zeta potential and particle sizes of the complexes were between 20mV and 50nm. COS-7 cells transfection and cytotoxicity results suggested that PEG-Polycarbam-SP had a high efficient ability of delivery DNA, also less cytotoxic than PEI 25kDa and Polycarbam-SP. Conclusion： PEG-Polycarbam-SP could be an efficient and less toxic non-viral gene delivery vector which has potential application in gene therapy.
出处
《现代生物医学进展》
CAS
2013年第30期5821-5824,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(30931881)
上海市闵行区2011年度技术创新项目-企校合作专项(2011MH093)
关键词
PEG修饰
精胺
基因载体
转染
毒性
PEG-modified
Spermine
Gene vector
Transfection
Cytotoxicity
