CATIONIC LIPOSOME-MEDIATED INTRAVENOUS GENE DELIVERY
CATIONIC LIPOSOME-MEDIATED INTRAVENOUS GENE DELIVERY
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DOI：
10.1074/jbc.270.42.24864
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发表时间：
1995-10-20
影响因子：
4.8
通讯作者：
DEBS, R
中科院分区：
文献类型：
--
作者：
LIU, Y;LIGGITT, D;DEBS, R
系统性基因转移为研究体内基因功能和基因调控以及人类基因治疗提供了新的机会。我们使用氯霉素乙酰转移酶报告基因，以检查几个参数的发展，有效的，阳离子脂质体介导的，静脉内（IV）小鼠基因转移的重要性。然后，我们证明了这种方法可以产生生物学上重要基因的高水平表达。具体而言，我们评估了表达载体设计与产生的系统基因表达水平的关系，并比较了单独静脉注射DNA与DNA脂质体复合物产生的转染水平。我们发现，异源内含子的位置和表达质粒中使用的启动子元件都显著影响产生的系统基因表达水平。虽然静脉注射质粒DNA单独转染每一个组织分析，脂质体介导的传递是更有效的。我们还确定了重复IV注射DNA脂质体复合物产生高水平的全身转染。第二次注射DNA脂质体复合物产生的基因表达水平至少与单次IV注射后一样高。因此，不像一些病毒载体，中和宿主免疫反应不限制重新表达，以下重新注射的DNA脂质体complexes.Finally，我们表明，表达载体，产生最高水平的氯霉素乙酰转移酶报告基因的表达也可以产生高水平的表达两个集落刺激因子基因在小鼠中。具体地说，IV注射脂质体复合到表达载体中，我们已经将鼠粒细胞-巨噬细胞-集落刺激因子cDNA或人粒细胞-CSF cDNA插入其中，产生相应集落刺激因子基因产物的循环水平，其与先前已经显示的生物学和治疗学上显著的水平相当。
Systemic gene transfer provides new opportunities for the analysis of gene function and gene regulation in vivo, as well as for human gene therapy. We used the chloramphenicol acetyltransferase reporter gene to examine several parameters important for the development of efficient, cationic liposome-mediated, intravenous (IV) gene transfer in mice. We then demonstrated that this approach can produce high level expression of biologically important genes. Specifically, we assessed the relationship of expression vector design to the level of systemic gene expression produced, and compared transfection levels produced by intravenously injecting DNA alone versus DNA liposome complexes. We found that both the position of the heterologous intron, and the promoter element used in the expression plasmid, significantly affected the level of systemic gene expression produced. Although intravenous injection of plasmid DNA alone transfected every tissue analyzed, liposome-mediated delivery was much more efficient. We also established that repeated IV injection of DNA liposome complexes produced high level systemic transfection. The second injection of DNA liposome complexes produced levels of gene expression at least as high as those following a single IV injection. Thus, unlike some viral vectors, a neutralizing host-immune response does not limit re-expression, following reinjection of DNA liposome complexes.Finally, we showed that the expression vectors which produced the highest levels of chloramphenicol acetyltransferase reporter gene expression could also produce high level expression of two colony stimulating factor genes in mice. Specifically, IV injection of liposomes complexed to expression vectors into which we had inserted either the murine granulocyte-macrophage-colony stimulating factor cDNA or the human granulocyte-CSF cDNA, produced circulating levels of the corresponding colony stimulating factor gene product comparable to levels which have been shown previously to be both biologically and therapeutically significant.