Enhancement of transfection efficiency using ligand-modified lipid vesicles

Jun You, Masamichi Kamihira, Shinji Iijima
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引用次数: 8

Abstract

Previously, we had developed a simple gene transfection technique for animal cells using cationic lipid vesicles; a commercially available synthetic cationic surfactant, dimethyldioctadecyl ammonium bromide (DDAB) was used for making lipid vesicles. In the present study, the lipid vesicles for receptor mediated gene transfer were modified with a ligand such as insulin and galactose residues to realize enhanced transfection efficiency and/or cell-specific gene transfection. The insulin-modified lipid vesicle solution mixed with the plasmid DNA (pCMVβ) was added to COS-7, NIH3T3, Hela or HepG2 cells; the transfection efficiency was increased 3–4-fold in all the cell lines tested. Furthermore, a mixture of the galactose-modified lipid vesicles and plasmid pCMVβ was added to HepG2 or HuH-6 cells expressing asialoglycoprotein receptors, and the transfection efficiency was increased 3–4-fold in these cell lines.

利用配体修饰脂质囊泡提高转染效率
此前,我们已经开发了一种简单的动物细胞基因转染技术,使用阳离子脂质囊泡;采用市售阳离子表面活性剂二甲基二十八烷基溴化铵(DDAB)制备脂质囊泡。在本研究中,利用胰岛素和半乳糖残基等配体修饰用于受体介导的基因转移的脂质囊泡,以提高转染效率和/或细胞特异性基因转染。将与质粒DNA (pCMVβ)混合的胰岛素修饰脂质泡溶液加入COS-7、NIH3T3、Hela或HepG2细胞;转染效率提高了3 - 4倍。此外,将半乳糖修饰的脂质囊泡和质粒pCMVβ的混合物加入到表达asialal糖蛋白受体的HepG2或HuH-6细胞中,转染效率提高了3 - 4倍。
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