Efficient gene delivery using anionic liposome-complexed polyplexes (LPDII)
Efficient gene delivery using anionic liposome-complexed polyplexes (LPDII)
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DOI：
10.1023/a:1010338219401
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发表时间：
2000-10-01
影响因子：
4
通讯作者：
Lee, RJ
中科院分区：
文献类型：
--
作者：
Guo, WJ;Lee, RJ
以阴离子脂质体复合复合物为载体的合成基因转移载体（LPDII载体）在培养的哺乳动物细胞中的转染效率较高。利用荧光素酶报告基因，系统评价了多阳离子与DNA比值、脂质与DNA比值、多阳离子选择和脂质组成对人口腔癌KB细胞的影响。对于含有聚l -赖氨酸和二酰基磷脂酰乙醇胺/半乙酰胆甾醇（DOPE/CHEMS）阴离子脂质体的LPDII配方，在恒定的脂质与DNA比下，多阳离子/DNA （N/P）比的增加导致转染活性的增加。同时，基因高效传递的最佳脂质与DNA比受氮磷比的影响，氮磷比越高，脂质与DNA比越高。对于DNA凝聚剂，可以用聚乙烯亚胺（PEI）代替聚赖氨酸作为配方中的DNA凝聚剂。对于脂质成分。CHEMS可以被油酸、磷酸二酯和磷脂酰丝氨酸等阴离子脂类替代，但DOPE不能。一种促梭性辅助脂质，不能被二油磷脂酰胆碱替代。与常用的阳离子脂质体或PET介导转染相比，LPDII制剂的细胞毒性显着降低，并且几种细胞系转染效率高。LPDII载体避免了有毒阳离子脂质的使用，在基因治疗中有潜在的应用前景。
Synthetic gene transfer vectors based on polyplexes complexed to anionic liposomes (LPDII vectors) were characterized for their transfection efficiency in cultured mammalian cells. The effects of polycation to DNA ratio, lipid to DNA ratio, choice of polycation and lipid composition were systematically evaluated in human oral carcinoma KB cells, using a luciferase reporter gene. For LPDII formulations containing poly-L-lysine and dioeoylphosphatidylethanolamine/cholesteryl hemisuccinate (DOPE/CHEMS) anionic liposomes, at a constant lipid to DNA ratio, an increase in the polycation/DNA (N/P) ratio resulted in an increase in transfection activity. Meanwhile, the optimal lipid to DNA ratio for efficient gene delivery was influenced by the N/P ratio used, and was increased at higher N/P ratios. For the DNA condensing agent, poly-L-lysine could be replaced by polyethylenimine (PEI) as the DNA condensing agent in the formulations. For the lipidic components. CHEMS could be replaced by other anionic lipids including oleic acid, dicetylphosphate and phosphatidylserine, but DOPE. a fusogenic helper lipid, could not be replaced by dioleolyphosphatidylcholine. LPDII formulation showed significantly less cytotoxicity compared to the commonly used cationic lipsomes or PET mediated transfection and several cell lines were transfected with high efficiency. LPDII vectors avoid the use of toxic cationic lipids and may have potential application in gene therapy.