Nonviral gene delivery to the lateral ventricles in rat brain: Initial evidence for widespread distribution and expression in the central nervous system
Nonviral gene delivery to the lateral ventricles in rat brain: Initial evidence for widespread distribution and expression in the central nervous system
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DOI：
10.1006/mthe.2001.0272
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发表时间：
2001-03-01
影响因子：
12.4
通讯作者：
Irion, VR
中科院分区：
文献类型：
--
作者：
Hecker, JG;Hall, LL;Irion, VR
使用 DNA 进行非病毒基因表达取决于几个因素。这些包括（i）递送和到达目标组织，（ii）防止细胞外降解，（iii）被感兴趣的细胞充分摄取，以及（iv）防止细胞内降解以允许翻译足够水平的细胞内或分泌蛋白。作为证明非病毒、阳离子脂质介导的基因治疗可行性的第一步，我们提供了热休克蛋白 Hsp70 和报告基因酶在注射到侧脑室后在大鼠中枢神经系统 (CNS) 中成功递送和表达的证据。基因传递是通过优化的质粒 DNA 配方完成的，该配方已与阳离子脂质 MLRI 复合。报告了编码绿色荧光蛋白 (GFP)、荧光素酶和 Hsp70 的 DNA 载体的结果。标准免疫荧光方法用于证明报告蛋白和 Hsp70 的广泛表达。已完成三个冠状切片的体视学分析，说明了表情沿纵轴的分布。这些初步发现支持进一步开发非病毒、脂质介导的基因传递技术，用于保护性细胞内蛋白的瞬时表达，并且代表了体内研究以确定潜在临床益处的重要一步。
The use of DNA for nonviral gene expression depends on several factors. These include (i) delivery and accessibility to the targeted tissue, (ii) protection from extracellular degradation, (iii) sufficient uptake by cells of interest, and (iv) protection from intracellular degradation to allow translation of adequate levels of intracellular or secreted proteins. As an initial step in demonstrating the feasibility of nonviral, cationic lipid-mediated gene therapy, we present evidence for the successful delivery and expression of heat shock protein Hsp70 and reporter gene enzymes in the central nervous system (CNS) of the rat after injection into the lateral ventricle. Gene delivery is accomplished using optimized formulations of plasmid DNA, which have been complexed with the cationic lipid MLRI. Results from DNA vectors encoding for green fluorescent protein (GFP), luciferase, and Hsp70 are reported. Standard immunofluorescent methods were used to demonstrate widespread expression of the reporter proteins and of Hsp70. Stereology analysis has been completed on three coronal sections, which illustrates the distribution of expression along the longitudinal axis. These initial findings support the further development of nonviral, lipid-mediated gene delivery technology for transient expression of protective, intracellular proteins and represent an important step leading to in vivo studies to identify potential clinical benefits.