Plasmid DNA size does not affect the physicochemical properties of lipoplexes but modulates gene transfer efficiency
Plasmid DNA size does not affect the physicochemical properties of lipoplexes but modulates gene transfer efficiency
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DOI：
10.1093/nar/27.19.3792
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发表时间：
1999-10-01
影响因子：
14.9
通讯作者：
Pitard, B
中科院分区：
文献类型：
--
作者：
Kreiss, P;Cameron, B;Pitard, B
基因治疗的临床应用主要依赖于高效基因转移载体的开发。大的DNA分子只能通过合成载体，如阳离子脂质和聚合物，转染到细胞中。因此，本研究旨在探索阳离子脂质dna颗粒的物理化学性质，质粒范围从900到52 500 bp。脂多胺胶束与不同长度的质粒DNA络合形成的脂丛，其胶体稳定性取决于电荷比，从而形成三个结构域，分别对应带负电荷、中性电荷和带正电荷的脂丛。脂质体的形态和结构由DNA和阳离子脂质的理化特性决定。因此，结构的片层间距由阳离子脂质决定，其球形形态由DNA决定。本研究的主要结果是脂多胺-DNA复合物的形态和结构特征不依赖于质粒DNA的长度。另一方面，夹在脂质双层之间的质粒DNA分子的数量影响了它们的基因转移能力。含有最短质粒DNA的脂多胺-DNA复合物是最有效的基因转移载体。
Clinical applications of gene therapy mainly depend on the development of efficient gene transfer vectors. Large DNA molecules can only be transfected into cells by using synthetic vectors such as cationic lipids and polymers. The present investigation was therefore designed to explore the physicochemical properties of cationic lipid-DNA particles, with plasmids ranging from 900 to 52 500 bp. The colloidal stability of the lipoplexes formed by complexing lipopolyamine micelles with plasmid DNA of various lengths, depending on the charge ratio, resulted in the formation of three domains, respectively corresponding to negatively, neutrally and positively charged lipoplexes. Lipoplex morphology and structure were determined by the physicochemical characteristics of the DNA and of the cationic lipid. Thus, the lamellar spacing of the structure was determined by the cationic lipid and its spherical morphology by the DNA. The main result of this study was that the morphological and structural features of the lipopolyamine-DNA complexes did not depend on plasmid DNA length. On the other hand, their gene transfer capacity was affected by the sire of plasmid DNA molecules which were sandwiched between the lipid bilayers. The most effective lipopolyamine-DNA complexes for gene transfer were those containing the shortest plasmid DNA.