Hepatocyte-targeted gene transfer by combination of vascularly delivered plasmid DNA and in vivo electroporation
Hepatocyte-targeted gene transfer by combination of vascularly delivered plasmid DNA and in vivo electroporation
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DOI：
10.1038/sj.gt.3302435
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发表时间：
2005-04-01
期刊：
影响因子：
5.1
通讯作者：
Hashida, M
中科院分区：
文献类型：
--
作者：
Sakai, M;Nishikawa, M;Hashida, M
为了增加肝中的转基因表达，在静脉内注射裸质粒DNA（pDNA）或用半乳糖基化聚（L-赖氨酸）或半乳糖基化聚乙烯亚胺制备的pDNA/肝靶向载体复合物后，向左外侧叶施加电脉冲。裸pDNA注射后的电穿孔（250 V/cm，5 ms/脉冲，12脉冲，4 Hz）使表达显著增加高达200 000倍;所获得的表达水平显著大于通过pDNA/载体复合物和电穿孔的组合所获得的表达水平。我们清楚地表明，当施加电脉冲时，表达依赖于pDNA的血浆浓度。肝细胞的分离揭示了裸pDNA的分布以及转基因表达在很大程度上对电穿孔叶中的肝细胞具有选择性。经血管内注射裸pDNA后再经电穿孔转染，表达转基因产物的细胞数显著增加（P
To increase transgene expression in the liver, electric pulses were applied to the left lateral lobe after intravenous injection of naked plasmid DNA (pDNA) or pDNA/liver targeting vector complex prepared with galactosylated poly(L-lysine) or galactosylated polyethyleneimine. Electroporation ( 250 V/cm, 5 ms/pulse, 12 pulses, 4 Hz) after naked pDNA injection dramatically increased the expression up to 200 000-fold; the expression level obtained was significantly greater than that achieved by the combination of pDNA/vector complex and electroporation. We clearly demonstrated that the expression was dependent on the plasma concentration of pDNA at the time when the electric pulses were applied. Separation of liver cells revealed that the distribution of naked pDNA as well as transgene expression was largely selective to hepatocytes in the electroporated lobe. The number of cells expressing transgene product using vascularly administered naked pDNA followed by electroporation was significantly (P