Dascalu A I, Ardeleanu R, Neamtu A, Maier S S, Uritu C M, Nicolescu A, Silion M, Peptanariu D, Calin M, Pinteala M
Centre of Advanced Research in Bionanoconjugates and Biopolymers, "Petru Poni" Institute of Macromolecular Chemistry, 700487 Iasi, Romania.
J Mater Chem B. 2017 Sep 14;5(34):7164-7174. doi: 10.1039/c7tb01722g. Epub 2017 Aug 21.
Efficient tools are still being searched for to substitute the viral vectors in nucleic acid delivery applications. One of the most severe constraints in producing them is related to the strict reproducibility of their molecular characteristics, which is ensured through the synthesis. In this work, we report an original route to obtain polycationic nanoentities with low variability, which are able to act as cooperating carriers for dsDNA complexation and transport. The carriers are synthesized by rigorous conjugation of β-cyclodextrin (β-CD) with precise ratios of 2 kDa branched poly(ethyleneimine) (b-PEI) and 0.75 kDa poly(ethylene glycol) (PEG). Low cytotoxicity was the key parameter of the carrier design, besides the highest possible transfection ability, and both of these features were proven by HeLa cell culture assays. A reporter gene which induces the expression of green fluorescent protein (GFP), inserted in a plasmid, was used to perform the necessary quantitative measurements. In silico molecular modelling guided the carrier design and confirmed the functional mimicry of histones in the tight and compact nucleosome-like spiral packaging of dsDNA. The carrier molecules, synthesized with high reproducibility, are expected to be feasible for application in gene transfection.
人们仍在寻找高效工具来替代核酸递送应用中的病毒载体。生产这些载体最严重的限制之一与它们分子特征的严格可重复性有关，而这种可重复性是通过合成来确保的。在这项工作中，我们报告了一种获得低变异性聚阳离子纳米实体的原始途径，这些纳米实体能够作为双链DNA络合和转运的协同载体。载体通过β-环糊精（β-CD）与精确比例的2 kDa支化聚（乙烯亚胺）（b-PEI）和0.75 kDa聚（乙二醇）（PEG）进行严格共轭合成。除了尽可能高的转染能力外，低细胞毒性是载体设计的关键参数，而这两个特征均通过HeLa细胞培养试验得到证实。插入质粒中的诱导绿色荧光蛋白（GFP）表达的报告基因用于进行必要的定量测量。计算机辅助分子建模指导了载体设计，并证实了在双链DNA紧密且紧凑的核小体样螺旋包装中组蛋白的功能模拟。以高重现性合成的载体分子有望在基因转染中得到应用。
