Kogure Kentaro, Moriguchi Rumiko, Sasaki Kentaro, Ueno Masaharu, Futaki Shiroh, Harashima Hideyoshi
CREST, Japan Science and Technology Agency (JST), Japan.
J Control Release. 2004 Aug 11;98(2):317-23. doi: 10.1016/j.jconrel.2004.04.024.
The development of a multifunctional envelope-type nano device (MEND) for use in a non-viral gene delivery system using a novel lipid film hydration method is described. This packaging method involves three steps: (i) DNA condensation with a polycation, (ii) lipid film hydration for the electrostatic binding of the condensed DNA, and (iii) sonication to package the condensed DNA with lipids. For DNA condensation, the optimum ratio of DNA to poly-L-lysine (PLL) was determined by titrating DNA and PLL. The condensed DNA/PLL complex (DPC) was coated with a lipid bilayer by lipid film hydration followed by sonication, which was confirmed by sucrose density gradient centrifugation. Packaging of DPC with lipids increased the transfection activity 10-fold over that of DPC. MEND, having octaarginine on the envelope as a device for membrane penetration to enhance cellular uptake, showed a 1000-fold higher transfection activity than DPC. The MEND developed in this study represents a promising non-viral gene delivery system.
本文描述了一种用于非病毒基因递送系统的多功能包膜型纳米装置（MEND）的开发，该装置采用了新型脂质膜水化方法。这种包装方法包括三个步骤：（i）用聚阳离子使DNA凝聚；（ii）脂质膜水化以实现凝聚DNA的静电结合；（iii）超声处理以将凝聚的DNA与脂质包装在一起。对于DNA凝聚，通过滴定DNA和聚-L-赖氨酸（PLL）来确定DNA与PLL的最佳比例。凝聚的DNA/PLL复合物（DPC）通过脂质膜水化然后超声处理被脂质双层包被，这通过蔗糖密度梯度离心得以证实。用脂质对DPC进行包装使转染活性比DPC提高了10倍。在包膜上具有八聚精氨酸作为膜穿透装置以增强细胞摄取的MEND，其转染活性比DPC高1000倍。本研究中开发的MEND代表了一种有前景的非病毒基因递送系统。
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