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首页> 外文期刊>Cell biology international. >Nuclear delivery of plasmid DNA determines the efficiency of gene expression
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Nuclear delivery of plasmid DNA determines the efficiency of gene expression

机译：质粒DNA的核递送决定了基因表达的效率

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Abstract As a cationic non‐viral gene delivery vector, poly(agmatine/ N , N ′‐cystamine‐bis‐acrylamide) (AGM‐CBA) showed significantly higher plasmid DNA (pDNA) transfection ability than polyethylenimine (PEI) in NIH/3T3 cells. The transfection expression of AGM‐CBA/pDNA polyplexes was found to have a non‐linear relationship with AGM‐CBA/pDNA weight ratios. To further investigate the mechanism involved in the transfection process of poly(AGM‐CBA), we used pGL3‐control luciferase reporter gene (pLUC) as a reporter pDNA in this study. The distribution of pLUC in NIH/3T3 cells and nuclei after AGM‐CBA/pLUC and PEI/pLUC transfection were determined by quantitative polymerase chain reaction (qPCR) analysis. The intracellular trafficking of the polyplexes was evaluated by cellular uptake and nuclei delivery of pLUC, and the intracellular availability was evaluated by the ratio of transfection expression to the numbers of pLUC delivered in nuclei. It was found that pLUC intracellular trafficking did not have any correlation with the transfection expression, while an excellent correlation was found between the nuclei pLUC availability and transfection expression. These results suggested that the intracellular availability of pLUC in nuclei was the rate‐limiting step for pLUC transfection expression. Further optimization of the non‐viral gene delivery system can be focused on the improvement of gene intracellular availability.
机译：摘要作为阳离子非病毒基因递送载体，聚（Agmatine / N，N'-cystamine-Bis-丙烯酰胺）（AGM-CBA）显示出明显高的质粒DNA（PDNA）转染能力，而不是NIH / 3T3中的聚乙烯（PEI）细胞。发现AGM-CBA / PDNA多种的转染表达与AGM-CBA / PDNA重量比具有非线性关系。为了进一步研究聚（AGM-CBA）的转染过程中涉及的机制，我们将PGL3-Contoil Luciferase报告基因（Pluc）用作本研究中的报告PDNA。通过定量聚合酶链反应（QPCR）分析测定AGM-CBA / pluc和PEI / PLUC转染后NIH / 3T3细胞和核核的分布。通过细胞吸收和核递送评估多种聚合物的细胞内运输，通过转染表达与核中递送的Pluc数量的比率来评价细胞内可用性。发现Pluc细胞内贩运没有与转染表达的任何相关性，而在核覆皮可用性和转染表达之间发现了优异的相关性。这些结果表明，核中胆量的细胞内可用性是Pluc转染表达的速率限制步骤。进一步优化非病毒基因递送系统可以集中于改善基因细胞内可用性。

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