Efficient synthesis and cell-transfection properties of a new multivalent cationic lipid for nonviral gene delivery
Efficient synthesis and cell-transfection properties of a new multivalent cationic lipid for nonviral gene delivery
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DOI：
10.1021/jm020233w
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发表时间：
2002-11-07
影响因子：
7.3
通讯作者：
Safinya, CR
中科院分区：
文献类型：
--
作者：
Ewert, K;Ahmad, A;Safinya, CR
脂质介导的DNA进入细胞的传递对于基因治疗和基础研究应用都有很大的希望。本文介绍了一种新的多价阳离子脂质与双支化的头基结构的基因传递应用的有效和简便的合成和表征。该合成方案可以扩展以得到不同电荷、间隔基或脂质链长度的阳离子脂质。阳离子脂质体（CL）与DNA的自组装复合物的化学和物理性质给出了为什么多价阳离子脂质具有上级转染性质的指示。该脂质在完全质子化状态下带有带有五个电荷的头基，其连接到基于3，4-二羟基苯甲酸的不饱和双链疏水部分。由新的多价脂质和中性脂质1，2-二油酰基-sn-甘油磷脂酰胆碱（DOPC）组成的脂质体用于制备与DNA的复合物。通过光学显微镜和小角X射线散射对这些复合物的结构进行的研究揭示了CL-DNA复合物的层状L-alpha（C）相，其中DNA分子夹在脂质的双层之间。使用含有萤火虫荧光素酶报告基因的质粒DNA的实验表明，这些复合物有效地抑制哺乳动物细胞。当与单价阳离子脂质2，3-二油烯基氧基丙基三甲基氯化铵（DOTAP）相比时，用新的多价脂质可实现的CL-DNA复合物的膜的较高电荷密度在阳离子与中性脂质的小摩尔比的方案中大大增加了转染效率。这是期望的，以使阳离子脂质的已知毒性作用最小化。
Lipid-mediated delivery of DNA into cells holds great promise both for gene therapy and basic research applications. This paper describes the efficient and facile synthesis and the characterization of a new multivalent cationic lipid with a double-branched headgroup structure for gene delivery applications. The synthetic scheme can be extended to give cationic lipids of different charge, spacer, or lipid chain length. The chemical and physical properties of self-assembled complexes of the cationic liposomes (CLs) with DNA give indications of why multivalent cationic lipids possess superior transfection properties. The lipid bears a headgroup with five charges in the fully protonated state, which is attached to an unsaturated double-chain hydrophobic moiety based on 3,4-dihydroxybenzoic acid. Liposomes consisting of the new multivalent lipid and the neutral lipid 1,2-dioleoyl-sn-glycerophosphatidylcholine (DOPC) were used to prepare complexes with DNA. Investigations of the structures of these complexes by optical microscopy and small-angle X-ray scattering reveal a lamellar L-alpha(C) phase of CL-DNA complexes with the DNA molecules sandwiched between bilayers of the lipids. Experiments using plasmid DNA containing the firefly luciferase reporter gene show that these complexes efficiently transfect mammalian cells. When compared to the monovalent cationic lipid 2,3-dioleyloxypropyltrimethylammonium chloride (DOTAP), the higher charge density of the membranes of CL-DNA complexes achievable with the new multivalent lipid greatly increases transfection efficiency in the regime of small molar ratios of cationic to neutral lipid. This is desired to minimize the known toxicity effects of cationic lipids.