School of Polymer Science and Engineering, The University of Southern Mississippi, Hattiesburg, Mississippi 39406-5050, United States.
School of Biological, Environmental, and Earth Sciences, The University of Southern Mississippi, Hattiesburg, Mississippi 39406-5050, United States.
Biomacromolecules. 2022 Jan 10;23(1):34-46. doi: 10.1021/acs.biomac.1c00824. Epub 2021 Nov 18.
The black-legged tick () is the primary vector for bacteria that cause Lyme disease (), where numerous glycosylated tick proteins are involved at the interface of vector-host-pathogen interactions. Reducing the expression of key tick proteins, such as selenoprotein K (SelK), through RNA interference is a promising approach to reduce pathogen transmission, but efficient delivery of nucleic acids to arthropods has proven challenging. While cationic glycopolymers have been used as nonviral gene delivery vehicles in mammalian cells, their use in arthropod or insect gene transfection has not been established. In this study, statistical acrylamide-based cationic glycopolymers with glucose or galactose pendant groups were synthesized by reversible addition-fragmentation chain transfer polymerization, and the effects of the saccharide pendant group and cationic monomer loading on polymer cytotoxicity, RNA complexation, and SelK gene knockdown in ISE6 cells were evaluated. All polymers exhibited low cytotoxicity, yet RNA/copolymer complex cell uptake and gene knockdown were highly dependent on the saccharide structure and the N:P (amino to phosphate groups) ratio.
黑腿蜱（）是引起莱姆病（）的细菌的主要传播媒介，在媒介-宿主-病原体相互作用的界面涉及许多糖基化蜱蛋白。通过 RNA 干扰降低关键蜱蛋白（如硒蛋白 K（SelK））的表达是减少病原体传播的一种很有前途的方法，但将核酸有效地递送到节肢动物一直具有挑战性。虽然阳离子糖聚合物已被用作哺乳动物细胞中的非病毒基因传递载体，但它们在节肢动物或昆虫基因转染中的应用尚未确定。在这项研究中，通过可逆加成-断裂链转移聚合合成了基于丙烯酰胺的统计阳离子糖聚合物，带有葡萄糖或半乳糖侧基，并评估了糖侧基和阳离子单体负载对聚合物细胞毒性、RNA 复合物形成以及 ISE6 细胞中 SelK 基因敲低的影响。所有聚合物均表现出低细胞毒性，但 RNA/聚合物复合物的细胞摄取和基因敲低高度依赖于糖的结构和 N：P（氨基与磷酸基团）比。
