Nasr Sarah S, Lee Sangeun, Thiyagarajan Durairaj, Boese Annette, Loretz Brigitta, Lehr Claus-Michael
Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University, 66123 Saarbrücken, Germany.
Department of Pharmacy, Saarland University, 66123 Saarbrücken, Germany.
Pharmaceutics. 2021 Nov 13;13(11):1924. doi: 10.3390/pharmaceutics13111924.
Co-delivery of different species of protein-encoding polynucleotides, e.g., messenger RNA (mRNA) and plasmid DNA (pDNA), using the same nanocarrier is an interesting topic that remains scarcely researched in the field of nucleic acid delivery. The current study hence aims to explore the possibility of the simultaneous delivery of mRNA (mCherry) and pDNA (pAmCyan) using a single nanocarrier. The latter is based on gelatin type A, a biocompatible, and biodegradable biopolymer of broad pharmaceutical application. A core-shell nanostructure is designed with a thermally stabilized gelatin-pDNA coacervate in its center. Thermal stabilization enhances the core's colloidal stability and pDNA shielding effect against nucleases as confirmed by nanoparticle tracking analysis and gel electrophoresis, respectively. The stabilized, pDNA-loaded core is coated with the cationic peptide protamine sulfate to enable additional surface-loading with mRNA. The dual-loaded core-shell system transfects murine dendritic cell line DC2.4 with both fluorescent reporter mRNA and pDNA simultaneously, showing a transfection efficiency of 61.4 ± 21.6% for mRNA and 37.6 ± 19.45% for pDNA, 48 h post-treatment, whereas established commercial, experimental, and clinical transfection reagents fail. Hence, the unique co-transfectional capacity and the negligible cytotoxicity of the reported system may hold prospects for vaccination among other downstream applications.
使用同一纳米载体共同递送不同种类的蛋白质编码多核苷酸，例如信使核糖核酸（mRNA）和质粒DNA（pDNA），是核酸递送领域中一个有趣但仍鲜少被研究的课题。因此，本研究旨在探索使用单一纳米载体同时递送mRNA（mCherry）和pDNA（pAmCyan）的可能性。后者基于A型明胶，这是一种具有广泛药物应用的生物相容性和可生物降解的生物聚合物。设计了一种核壳纳米结构，其中心为热稳定的明胶-pDNA凝聚层。热稳定性分别通过纳米颗粒跟踪分析和凝胶电泳得到证实，增强了核心的胶体稳定性和pDNA对核酸酶的屏蔽作用。稳定的、负载pDNA的核心用阳离子肽硫酸鱼精蛋白包被，以便能够额外在表面负载mRNA。双负载核壳系统能够同时用荧光报告mRNA和pDNA转染小鼠树突状细胞系DC2.4，在处理后48小时，mRNA的转染效率为61.4±21.6%，pDNA的转染效率为37.6±19.45%，而现有的商业、实验和临床转染试剂则无法做到。因此，所报道系统独特的共转染能力和可忽略不计的细胞毒性在其他下游应用中，可能为疫苗接种带来前景。
