Department of Medicine, Division of Infectious Diseases, Section of Virology , Imperial College London , Norfolk Place, London W21PG , U.K.
Polymer Chemistry Laboratory, School of Engineering and Materials Science , Queen Mary University of London , London E1 4NS , U.K.
Biomacromolecules. 2018 Jul 9;19(7):2870-2879. doi: 10.1021/acs.biomac.8b00429. Epub 2018 May 3.
Nucleic acid delivery systems are commonly translated between different modalities, such as DNA and RNA of varying length and structure, despite physical differences in these molecules that yield disparate delivery efficiency with the same system. Here, we synthesized a library of poly(2-ethyl-2-oxazoline)/poly(ethylene imine) copolymers with varying molar mass and charge densities in order to probe how pDNA, mRNA, and RepRNA polyplex characteristics affect transfection efficiency. The library was utilized in a full factorial design of experiment (DoE) screening, with outputs of luciferase expression, particle size, surface charge, and particle concentration. The optimal copolymer molar mass and charge density was found as 83 kDa/100%, 72 kDa/100%, and 45 kDa/80% for pDNA, RepRNA, and mRNA, respectively. While 10 of the synthesized copolymers enhanced the transfection efficiency of pDNA and mRNA, only 2 copolymers enhanced RepRNA transfection efficiency, indicating a narrow and more stringent design space for RepRNA. These findings suggest that there is not a "one size fits all" polymer for different nucleic acid species.
核酸传递系统通常在不同的模式之间进行转换，例如不同长度和结构的 DNA 和 RNA，尽管这些分子在物理上存在差异，但它们在相同的系统中表现出不同的传递效率。在这里，我们合成了一系列具有不同摩尔质量和电荷密度的聚（2-乙基-2-恶唑啉）/聚（聚乙烯亚胺）共聚物，以探究 pDNA、mRNA 和 RepRNA 多聚物的特性如何影响转染效率。该库用于全因子设计实验（DoE）筛选，输出为荧光素酶表达、粒径、表面电荷和颗粒浓度。对于 pDNA、RepRNA 和 mRNA，发现最佳共聚物摩尔质量和电荷密度分别为 83 kDa/100%、72 kDa/100%和 45 kDa/80%。虽然 10 种合成的共聚物增强了 pDNA 和 mRNA 的转染效率，但只有 2 种共聚物增强了 RepRNA 的转染效率，表明 RepRNA 的设计空间更窄且更严格。这些发现表明，对于不同的核酸种类，没有一种“一刀切”的聚合物。
