Coulberson Arlena L, Hud Nicholas V, LeDoux Joseph M, Vilfan Igor D, Prausnitz Mark R
School of Chemical Engineering, Georgia Institute of Technology, Atlanta 30332-0100, USA.
J Control Release. 2003 Jan 17;86(2-3):361-70. doi: 10.1016/s0168-3659(02)00417-0.
To develop improved methods of gene delivery, packaging DNA in chemical or viral vectors could increase electroporation-mediated transfection. To test this hypothesis, electroporation was applied to DU145 prostate cancer cells incubated with green fluorescent protein-encoded DNA plasmid either naked or packaged with cationic lipid (Lipofectin), polycationic peptide (salmon protamine) or retroviral vectors (Moloney murine leukemia viruses) and then assayed for gene expression and cell viability. Cationic lipid or electroporation alone each significantly increased transfection, but their combination was less effective. Addition of protamine peptide during electroporation was also less effective than electroporation alone. The combination of retroviral vectors and electroporation transfected fewer cells than retrovirus alone. We conclude that the combination of electroporation with chemical or viral vectors does not improve gene transfection in vitro.
为了开发改进的基因递送方法,将DNA包装在化学或病毒载体中可提高电穿孔介导的转染效率。为了验证这一假设,将电穿孔应用于与绿色荧光蛋白编码的DNA质粒一起孵育的DU145前列腺癌细胞,该质粒可以是裸质粒,也可以用阳离子脂质(Lipofectin)、聚阳离子肽(鲑鱼精蛋白)或逆转录病毒载体(莫洛尼鼠白血病病毒)包装,然后检测基因表达和细胞活力。单独使用阳离子脂质或电穿孔均可显著提高转染效率,但两者联合使用效果较差。在电穿孔过程中添加精蛋白肽的效果也不如单独使用电穿孔。逆转录病毒载体与电穿孔联合使用转染的细胞比单独使用逆转录病毒转染的细胞少。我们得出结论,电穿孔与化学或病毒载体联合使用并不能提高体外基因转染效率。