A DNA delivery system containing listeriolys in Oresults in enhanced hepatocytedirected gene expression

作     者：Cherie M. Walton, Catherine H. Wu and George Y. Wu 

作者机构：Department of Medicine Division of Gastroenterology-Hepatology University of Connecticut Health Center Farmington CT United States Department of Medicine Division of Gastroenterology-Hepatology University of Connecticut Health Center Farmington CT 06030 United States 

出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)

年 卷 期：1999年第5卷第6期

页      面：465-469页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学] 

基　　金：supported in part by agrant from the National Institutes of He alth,DK-42182(GYW) the Immune Response Cor por ation(GHW) the Herman Lopata Chair for He patitis Research 

主　　题：DNA gene expression listeriolysin O hepatocytes 

摘      要：AIM To determine whether incorporation of the pH dependent bacterial toxin listeriolysin O (LLO) into the DNA carrier system could increase the endosomal escape of internalized DNA and result gene expression. METHODS A multi component delivery system was prepared consisting of asialoglycoprotein (ASG), poly L lysine (PL), and LLO. Two marker genes, luciferase and β galactosidase in plasmids were complexed and administered in vitro to Huh7[ASG receptor (+) ] and SK Hep1[ASG receptor (-) ] cells. Purity, hemolytic activity, gene expression, specificity, and toxicity were evaluated. RESULTS An LLO containing conjugate retained cell targeting specificity and membranolytic activity. In ASG receptor (+) cells, luciferase gene expression was enhanced by more than 7 fold over that of conjugates without the incorporation of listeriolysin O. No significant expression occurred in ASG receptor (-) cells. Enhancement of β galactosidase gene expression was less, but still significantly increased over controls. There was no detectable toxicity at concentrations shown to be effective in transfection studies. CONCLUSIONS ASOR PL can be coupled to LLO using disulfide bonds, and successfully target and increase the gene expression of foreign DNA.