Transition from a normal to inverted cylinder for an amidine-bearing lipid/pDNA complex and its excellent transfection.
Transition from a normal to inverted cylinder for an amidine-bearing lipid/pDNA complex and its excellent transfection.
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DOI：
10.1021/bc050164a
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发表时间：
2005-10
影响因子：
4.7
通讯作者：
Kazunori Koiwai;K. Tokuhisa;Ryouji Karinaga;Y. Kudo;S. Kusuki;Y. Takeda;K. Sakurai
中科院分区：
文献类型：
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作者：
Kazunori Koiwai;K. Tokuhisa;Ryouji Karinaga;Y. Kudo;S. Kusuki;Y. Takeda;K. Sakurai
合成了一种含酰胺基的阳离子脂类(TRX)，并将其与另外两种中性磷脂混合，制得脂复合体(即质粒DNA+脂质复合体)。小角X射线散射表明，DNA的加入导致了从正常六角填充圆柱体到倒置六角填充圆柱体的结构转变。透射电子显微镜显示，加入DNA后，线状胶束转变为直径约50-200 nm的球形胶束。结合这些结果得出结论，复杂的DNA是六角形的堆积(或浓缩)成球形聚集体。其大小和形态被认为适合于内吞或囊泡融合。将pDNA(pEGFP-C1)与Trx形成的复合体转入Hep G2细胞。流式细胞仪和共聚焦显微镜显示，该系统比传统的转染剂表达更多的绿色荧光蛋白(GFP)。此外，TRX的细胞毒性低于其他转染剂。本文提出了烷基作为基因转染器的可能性。
A cationic lipid (TRX) having an amidine headgroup was synthesized, and a lipoplex (i.e., plasmid DNA+lipid complex) was prepared from the mixture of TRX and two other neutral colipids. Small-angle X-ray scattering showed that the addition of DNA induced a structural transition from normal to inverted hexagonally packed cylinders. Transmission electron microscopy showed a threadlike micelle was transformed to a spherical micelle about 50-200 nm in diameter by adding DNA. A combination of these results leads to the conclusion that the complexed DNA is hexagonally packed (or condensed) into spherical aggregates. The size and morphology are believed suitable for endocytosis uptake or vesicle fusion. The complex made from pDNA (pEGFP-C1) and TRX was transfected to Hep G2. Flow cytometry and confocal microscopy showed that the present system expressed green fluorescence protein (GFP) more than a conventional transfection reagent. Additionally, TRX was less cytotoxic than other transfection reagents. This paper presents the attractive possibility of the amidine group for a transfection device.