Mizuguchi H, Nakagawa T, Morioka Y, Imazu S, Nakanishi M, Kondo T, Hayakawa T, Mayumi T
Faculty of Pharmaceutical Science, Osaka University, Japan.
Biochem Biophys Res Commun. 1997 May 8;234(1):15-8. doi: 10.1006/bbrc.1997.6568.
Development of methodologies for gene transfer into the central nervous system (CNS) is important for fundamental research as well as clinical studies for gene therapy. Cationic liposomes (CL) are attractive vectors because of their safety and ease of use. However, to date only low rates of success have been reported. We succeeded in obtaining high transfection efficiencies into the newborn mouse brain in vivo by CL and a cytoplasmic gene expression system based on T7 RNA polymerase and T7 RNA polymerase- and the luciferase-gene with the T7 promoter sequence. This system showed an efficiency rate 2 orders of magnitude higher than the standard system, which used CL and luciferase genes with a Rous sarcoma virus promoter, pRSVL. In addition, in vitro experiments using LLCMK2 cells showed that cytoplasmic gene expression occurred rapidly (within 6 h) after transfection. In contrast, pRSVL required 24-48 h for induction of luciferase expression. Our results suggest that the cytoplasmic gene expression system is useful for gene delivery into the CNS.
开发将基因导入中枢神经系统（CNS）的方法对于基础研究以及基因治疗的临床研究都很重要。阳离子脂质体（CL）因其安全性和易用性而成为有吸引力的载体。然而，迄今为止报道的成功率很低。我们通过CL以及基于T7 RNA聚合酶和T7 RNA聚合酶的细胞质基因表达系统，成功地在新生小鼠脑内实现了高转染效率，该系统带有T7启动子序列的荧光素酶基因。该系统显示出的效率比使用CL和带有劳氏肉瘤病毒启动子pRSVL的荧光素酶基因的标准系统高2个数量级。此外，使用LLCMK2细胞进行的体外实验表明，转染后细胞质基因表达迅速发生（6小时内）。相比之下，pRSVL诱导荧光素酶表达需要24 - 48小时。我们的结果表明，细胞质基因表达系统对于将基因递送至中枢神经系统是有用的。
