Abstract

It is very challenging to fabricate a non-viral gene carrier system containing multifunctional components to overcome multiple extra- and intracellular barriers. Herein, we developed a novel PEGylated, targeted and fluorescence traceable multifunctional supramolecular host-guest gene vector system. This system was based on poly(β-cyclodextrin) (PCD), disulfide-containing azobenzene-terminated branched polymer (Az-ss-BPDM-RhB) with rhodamine-fluorescent functional groups and adamantyl terminated folate-modified PEG (Ad-PEG-FA). Transfection and cellular uptake data of the PEGylated supramolecular PCD/Az-ss-BPDM-RhB/DNA polyplexes demonstrated that prolonged incubation time to 24 h or longer could improve cell uptake and transfection efficiency of the PEGylated polyplexes. Good targeting and fluorescence traceable feature of this host-guest interaction gene delivery system was displayed. Moreover, the PEGylated and targeted supramolecular PCD/Az-ss-BPDM-RhB/Ad-PEG-FA showed lower cytotoxicity and in vivo zebrafish embryo toxicity and better gene transfection efficiency compared with the control 25 kDa PEI. Therefore, this PEGylated and targeted multifunctional supramolecular platform is a promising and effective gene delivery system for fluorescence traceable applications.

摘要

构建包含多功能组件的非病毒基因载体系统以克服多种细胞外和细胞内屏障是非常具有挑战性的。在此，我们开发了一种新型聚乙二醇化、靶向和荧光可追踪的多功能超分子主客体基因载体系统。该系统基于聚（β-环糊精）（PCD）、含二硫化物的偶氮苯封端支化聚合物（Az-ss-BPDM-RhB）和罗丹明荧光官能团和金刚烷基封端的叶酸修饰 PEG（Ad-PEG- F A）。PEG 化超分子 PCD/Az-ss-BPDM-RhB/DNA 复合物的转染和细胞摄取数据表明，将孵育时间延长至 24 小时或更长时间可以提高 PEG 化复合物的细胞摄取和转染效率。展示了该主客体相互作用基因递送系统良好的靶向性和荧光可溯源性。此外，聚乙二醇化和靶向超分子 PCD/Az-ss-BPDM-RhB/Ad-PEG-FA 显示出较低的细胞毒性和与对照 25 kDa PEI 相比，体内斑马鱼胚胎毒性和更好的基因转染效率。因此，这种聚乙二醇化和靶向多功能超分子平台是一种用于荧光可溯源应用的有前途且有效的基因传递系统。