Characterization of cationic liposome-mediated gene transfer in vivo by intravenous administration
Characterization of cationic liposome-mediated gene transfer in vivo by intravenous administration
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DOI：
10.1089/hum.1997.8.13-1585
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发表时间：
1997-09-01
影响因子：
4.2
通讯作者：
Liu, DX
中科院分区：
文献类型：
--
作者：
Song, YK;Liu, F;Liu, DX
研究了阳离子脂质体的理化性质，包括阳离子脂类的结构、阳离子脂类与DNA的比例、脂质体的颗粒大小和辅助脂类的包裹体对体内基因表达水平、位置和持续时间的影响。利用含有荧光素酶或绿色荧光蛋白基因的巨细胞病毒(CMV)驱动的基因表达系统，以及两种阳离子脂质[N-(2，3-二油酰氧基)丙基-N，N，N-三甲基氯化铵(DOTMA)和1，2-二油酰氧基-3-三甲基丙烷(DOTAP)]，我们在体内通过一次静脉注射DNA/脂质体复合物的方法证明了阳离子脂质体能够在小鼠的肺、心、肝、脾和肾脏等器官中导入细胞。转染率主要由阳离子脂的结构和阳离子脂与DNA的比例决定。尽管DOTAP脂质体中胆固醇的存在不影响转染率，但DOTAP或DOTMA脂质体中的二油酰磷脂酰乙醇胺(DOPE)显著降低了脂质体的体内转染率。从时间进程来看，基因在不同器官中的表达是短暂的，峰值在4~24小时，到第4天下降到峰值的不到1%。重复注射的实验表明，基因表达的峰值可以在随后的注射中恢复。
Physicochemical properties of the cationic liposomes, including structure of the cationic lipids, cationic lipid-to-DNA ratio, liposome particle size, and inclusion of the helper lipids, were studied for their effect on the level, site, and duration time of gene expression in vivo by intravenous administration. Using a cytomegalovirus (CMV)-driven gene expression system containing either the luciferase or green fluorescence protein gene as a reporter and two cationic lipids [N-(2,3-dioleoyloxy)propyl-N,N,N-trimethylammonium chloride (DOTMA) and 1,2-dioleoyloxy-3-trimethylammonium propane (DOTAP)], we demonstrated in vivo by a single intravenous injection of DNA/liposome complexes into mice, that cationic liposomes are capable of transfecting cells in organs such as the lung, heart, liver, spleen, and kidney. Transfection efficiency is determined mainly by the structure of the cationic lipid and the ratio of cationic lipid to DNA. Although the presence of cholesterol in DOTAP liposomes did not affect transfection activity, inclusion of dioleoylphosphatidylethanolamine (DOPE) into either DOTAP or DOTMA liposomes significantly decreases liposome transfection activity in vivo. Results from time course show that gene expression in different organs is transient, with a peak level between 4 and 24 hr, dropping to less than 1% of the peak level by day 4. Experiments with repeated injections showed that the peak level of gene expression could be regained by subsequent injection.