Preparation of efficient gene carriers using a polyamidoamine dendron-bearing lipid: Improvement of serum resistance
Preparation of efficient gene carriers using a polyamidoamine dendron-bearing lipid: Improvement of serum resistance
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DOI：
10.1021/bc050012f
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发表时间：
2005-09-01
影响因子：
4.7
通讯作者：
Kono, K
中科院分区：
文献类型：
--
作者：
Takahashi, T;Harada, A;Kono, K
在以前的研究中，我们开发了一种新的阳离子脂质组成的第三代和两个十二烷基链的聚酰胺胺树枝状分子，命名为DL-G3，它与fasogenic脂质二油酰磷脂酰乙醇胺（DOPE）的组合实现了有效的转染CV1细胞的质子海绵效应和膜融合的协同作用。本研究检测了血清对DL-G3 -DOPE -质粒DNA脂质复合物的转染活性的影响。具有在不存在血清的情况下优化的组合物的脂质复合物的转染活性在存在血清的情况下显著降低。然而，在血清存在下诱导有效转染的脂质复合物可通过控制DL-G3的伯胺与磷酸基团的电荷比（N/P比）和DOPE含量来获得。该复合物在有血清存在下表现出最高的转染活性，具有较低的N/P比和较高的DOPE含量。而这些复合物的解体诱导加入肝素，这是一种带负电荷的基团的多糖，复合物，保留转染活性的血清的存在下，需要更多的负电荷的肝素复合物解体。该结果意味着其对带负电荷的血清蛋白的更高稳定性。将抗血清复合物与一些市售转染试剂（例如Lipofectamine和SuperFect）进行比较，表明DL-G3复合物在血清存在下实现了对这些细胞更有效的转染。
In a previous study, we developed a novel cationic lipid consisting of polyamidoamine dendron of third generation and two dodecyl chains, designated as DL-G3, which in combination with a fasogenic lipid dioleoylphosphatidylethanolamine (DOPE) achieves efficient transfection of CV1 cells by synergetic action of the proton sponge effect and membrane fusion. This study examines the effect of serum on the transfection activity of the DL-G3 -DOPE -plasmid DNA lipoplexes. The transfection activity of a lipoplex with a composition optimized in the absence of serum decreased markedly in the presence of serum. However, the lipoplexes that induce efficient transfection in the presence of serum were obtainable by controlling the charge ratio of the primary amine of the DL-G3 to the phosphate group (N/P ratio) and DOPE content. The complex, which exhibited the highest transfection activity in the presence of serum, has a lower N/P ratio and higher DOPE content than that optimized in the absence of serum. Whereas disintegration of these complexes was induced by addition of heparin, which is a polysaccharide with negatively charged groups, the complex that retained transfection activity in the presence of serum required more negative charges of heparin for complex disintegration. That result implies its higher stability against negatively charged serum proteins. Comparison of the serum-resistant complex with some commercially available transfection reagents, such as Lipofectamine and SuperFect, indicates that the DL-G3 complex achieved more efficient transfection of these cells in the presence of serum.