Evaluation of cationic liposome suitable for gene transfer into pregnant animals
Evaluation of cationic liposome suitable for gene transfer into pregnant animals
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DOI：
10.1006/bbrc.1999.0590
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发表时间：
1999-05-10
影响因子：
3.1
通讯作者：
Terada, M
中科院分区：
文献类型：
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作者：
Ochiya, T;Takahama, Y;Terada, M
阳离子脂质体介导的体内基因转移是一种很有前途的体细胞基因治疗方法。为了评估最适合的脂质体基因传递到广泛的器官和胎儿的小鼠，我们已经探索了几种类型的阳离子脂质体共轭质粒DNA携带β-半乳糖苷酶基因通过静脉注射到怀孕的动物。通过Southern印迹分析评估转导效率，并通过β-半乳糖苷酶活性的酶促证明评估转移基因的表达。通过分析几种类型的最近合成的阳离子脂质体/脂质制剂，DMRIE-C试剂，阳离子脂质DMRIE（1，2-二肉豆蔻氧基丙基-3-二甲基-羟乙基溴化铵）和胆固醇在膜过滤水中的脂质体制剂符合我们的要求。当在交配后第11.5天（p.c.）将质粒DNA/DMRIE-C复合物静脉内施用到怀孕小鼠中时，在母鼠的几个器官中观察到转移的基因并表达。此外，虽然转移到胚胎中的拷贝数很低，但我们观察到子代中的报告基因表达。(C)北京：科学出版社.
Cationic liposome-mediated in vivo gene transfer represents a promising approach for somatic gene therapy. To assess the most suitable liposome for gene delivery into a wide range of organs and fetuses in mice, we have explored several types of cationic liposomes conjugated with plasmid DNA carrying the beta-galactosidase gene through intravenous injection into pregnant animals. Transduction efficiency was assessed by Southern blot analysis and expression of the transferred gene was evaluated by enzymatic demonstration of beta-galactosidase activity. Through the analysis of several types of recently synthesized cationic liposome/lipid formulations, DMRIE-C reagent, a liposome formulation of the cationic lipid DMRIE (1,2-dimyristyloxypropyl-3-dimethyl-hydroxy ethyl ammonium bromide) and cholesterol in membrane-filtered water met our requirements. When the plasmid DNA/DMRIE-C complexes were administered intravenously into pregnant mice at day 11.5 post coitus (p.c.), transferred genes were observed in several organs in dams and were expressed. Furthermore, although the copy numbers transferred into embryos were low, we observed reporter gene expression in the progeny. (C) 1999 Academic Press.