Ferrari Marilyn E, Rusalov Denis, Enas Joel, Wheeler Carl J
Department of Chemistry, Vical Incorporated, 9373 Towne Centre Drive, San Diego, CA 92121, USA.
Nucleic Acids Res. 2002 Apr 15;30(8):1808-16. doi: 10.1093/nar/30.8.1808.
The large number of cytofectin and co-lipid combinations currently used for lipoplex-mediated gene delivery reflects the fact that the optimal cytofectin/co-lipid combination varies with the application. The effects of structural changes in both cytofectin and co-lipid were systematically examined to identify structure-activity relationships. Specifically, alkyl chain length, degree of unsaturation and the head group to which the alkyl side chain was attached were examined to determine their effect on lipoplex structure and biological activity. The macroscopic lipoplex structure was assessed using a dye-binding assay and the biological activity was examined using in vitro transfection in three diverse cell lines. Lipoplexes were formulated in three different vehicles currently in use for in vivo delivery of naked plasmid DNA (pDNA) and lipoplex formulations. The changes in dye accessibility were consistent with structural changes in the lipoplex, which correlated with alterations in the formulation. In contrast, transfection activity of different lipoplexes was cell type and vehicle dependent and did not correlate with dye accessibility. Overall, the results show a correlation between transfection and enhanced membrane fluidity in both the lipoplex and cellular membranes.
目前用于脂质体介导的基因递送的大量细胞转染试剂和共脂质组合反映了这样一个事实，即最佳的细胞转染试剂/共脂质组合会因应用而异。系统地研究了细胞转染试剂和共脂质结构变化的影响，以确定构效关系。具体而言，研究了烷基链长度、不饱和度以及连接烷基侧链的头部基团，以确定它们对脂质体结构和生物活性的影响。使用染料结合测定法评估宏观脂质体结构，并在三种不同的细胞系中通过体外转染检测生物活性。脂质体在目前用于体内递送裸质粒DNA（pDNA）和脂质体制剂的三种不同载体中配制。染料可及性的变化与脂质体的结构变化一致，这与制剂的改变相关。相比之下，不同脂质体的转染活性取决于细胞类型和载体，与染料可及性无关。总体而言，结果表明脂质体和细胞膜中的转染与增强的膜流动性之间存在相关性。
