A smart nanoassembly consisting of acid-labile vinyl ether PEG-DOPE and protamine for gene delivery: preparation and in vitro transfection.
A smart nanoassembly consisting of acid-labile vinyl ether PEG-DOPE and protamine for gene delivery: preparation and in vitro transfection.
复制标题
DOI：
10.1021/bm800706f
复制
发表时间：
2008-10
影响因子：
6.2
通讯作者：
Zhenghong Xu;Wangwen Gu;Lingli Chen;Yu Gao;Zhiwen Zhang;Yaping Li
中科院分区：
文献类型：
--
作者：
Zhenghong Xu;Wangwen Gu;Lingli Chen;Yu Gao;Zhiwen Zhang;Yaping Li
提出了一种模块化设计的病毒样非病毒载体的概念，用于基因传递。最近，我们构建了一种具有多功能组分的新型智能纳米组件（SNA），该组件由含有鱼精蛋白硫酸盐（PS）的pDNA凝聚核和含有聚乙二醇-乙烯醚-涂料（PVD）的二油基磷脂酰乙醇胺（DOPE）脂质包膜组成。采用脂膜水合技术制备了mPEG 2000 （SNAs1）或mPEG 5000 （SNAs2）负载PS/DNA的SNAs。负载PS/DNA （10:1 w/w）的SNAs1和SNAs2的粒径分别约为160 nm和240 nm， zeta电位约为4 mV。体外释放实验表明，PVD具有良好的自脱pegylation能力，可使DOPE在低ph下恢复良好的促融合能力。SNAs对HEK 293、HeLa和COS-7细胞的转染效率比Lipofectamine 2000高，细胞毒性远低于Lipofectamine 2000。细胞摄取和亚细胞定位表明，SNAs的优良转染效率可能是由于含有PVD的dope脂质膜增加了细胞质中的PS/DNA，而鱼精蛋白增强了核传递或克服了核膜屏障。这些结果表明，基于pvd的纳米组件装载PS/DNA可能是一种很有前途的基因传递系统。
The conception of a modular designed and viruslike nonviral vector has been presented for gene delivery. Recently, we constructed a new smart nanoassembly (SNA) with multifunctional components that was composed of a condensed core of pDNA with protamine sulfate (PS) and a dioleoyl phosphatidylethanolamine (DOPE)-based lipid envelope containing poly(ethylene glycol)--vinyl ether--DOPE (PVD). SNAs with mPEG 2000 (SNAs1) or mPEG 5000 (SNAs2) loading PS/DNA were prepared by the lipid film hydration technique. The particle size was about 160 nm for SNAs1 and 240 nm for SNAs2 loading PS/DNA (10:1 w/w), and the zeta potential was about 4 mV for two SNAs. The in vitro release experiment indicated that PVD possessed a good ability for self-dePEGylation, which could result in the recovery of an excellent fusogenic capacity of DOPE at low pH. SNAs showed a higher transfection efficiency and much lower cytotoxicity than did Lipofectamine 2000 on HEK 293, HeLa, and COS-7 cells. The cellular uptake and subcellular localization demonstrated that the superior transfection efficiency of SNAs could result from the fact that the DOPE-based lipid envelope containing PVD increased PS/DNA in the cytoplasm, and protamine enhanced the nuclear delivery or overcame the nuclear membrane barrier. These results implied that the PVD-based nanoassembly loading PS/DNA could be a promising gene delivery system.