鼠源树突状细胞中鱼精蛋白和轮蛋白包埋脂质纳米粒子的转基因表达比较研究

Sharif M. Shaheen, A.K. Azad, M. Mustafezur Rahman, Md. Jashim Uddin
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引用次数: 0

摘要

基因治疗依赖于完美的DNA递送到细胞核亚结构域,在那里,DNA应该在基于脂质的程序化包装中进行协调,如多功能包膜型纳米器件(MEND)和其他地方发表的四片层多功能包膜类型纳米器件(T‐MEND)。在这两种包装系统中,DNA都必须与阳离子聚合物形成复合物(核心)。重要的是,程序包装内的这种DNA/聚集体核心的作用如何反映转基因表达。在这里,我们比较了包装在MEND中的精蛋白复合体(精蛋白-DNA)和轮蛋白复合体(聚轮烷-DNA)中萤火虫萤光素酶基因的转基因表达。用这些纳米颗粒转染树突状细胞,并测量转基因表达、细胞活力以及抗原呈递。在JAWs-II细胞(树突状细胞系)和骨髓树突状细胞中,来自前体MEND的转基因表达显著较高。当比较含有鱼精蛋白和轮状蛋白的T‐MEND包装系统时,鱼精蛋白的细胞活力和抗原呈递也比多聚蛋白迅速且更好。荧光标记的质粒DNA的共聚焦显微镜研究反映了DNA从前体复合体比轮状复合体更容易去凝聚的证据。我们的结果表明,天然的DNA冷凝器(鱼精蛋白复合体中的鱼精蛋白)比合成的聚阳离子冷凝器(轮状复合体中的聚轮烷)更喜欢转基因表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A comparative transgene expression study between a protaplex and a rotaplex embedded lipid-nano particles in murine derived dendritic cell

A comparative transgene expression study between a protaplex and a rotaplex embedded lipid-nano particles in murine derived dendritic cell

Gene therapy depends on the perfect DNA delivery to the nuclear subdomain, where DNA should be orchestrated in a lipid based programmed packaging, like multi-functional envelope type of nano device (MEND) and tetra lamellar multi-functional envelope type of nano device (T-MEND) published elsewhere. In both the packaging system, DNA has to make a complex (core) with a cationic polymer. It is important how about the effect of such a DNA/condenser core inside the programmed-packaging reflects the transgene expression. Here we compared transgene expression of a firefly luciferase gene from both the protaplex (protamine-DNA) and rotaplex (polyrotaxane-DNA), packaged in MENDs. Dendritic cells were transfected with these nanoparticles and transgene expression, cell viability as well as antigen presentations were measured. Transgene expression from the MEND of protaplex was significantly higher in JAWs-II cell (dendritic cell line) and bone marrow dendritic cell. Cell viability and antigen presentations from the protaplex were also prompt and better than that of polyplex, when T-MEND package systems containing protaplex and rotaplex, were compared. Confocal microscopic studies of fluorescently labeled plasmid DNA reflected the evidence of ease decondensation of DNA from the protaplex than that of a rotaplex. Our results suggest that a natural DNA condenser (protamine in protaplex) prefers transgene expression better to those of synthetic polycationic condenser (polyrotaxane in rotaplex).

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