Characteristics of transcription-regulatory elements for gene expression from plasmid vectors in human trophoblast cell lines
Characteristics of transcription-regulatory elements for gene expression from plasmid vectors in human trophoblast cell lines
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DOI：
10.1016/j.placenta.2006.02.007
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发表时间：
2006-09-01
期刊：
影响因子：
3.8
通讯作者：
Watanabe, Y.
中科院分区：
文献类型：
--
作者：
Komiya, E.;Kondoh, M.;Watanabe, Y.
非病毒基因传递系统可用于滋养层的基础研究。在这些系统中，基因表达受到质粒内调控元件盒的调控，并且转录活性在细胞系之间存在差异。在本研究中，我们使用 BeWo 和 JAR 人类滋养层细胞系系统地比较了几种表达盒和由猿病毒 40 (SV40) 启动子、多腺苷酸化 (PA) 信号和增强子组成的对照质粒的转录活性。我们还发现内含子元件的插入按以下顺序增强转录活性：内含子A>杂合β-珠蛋白-免疫球蛋白内含子>无内含子。在测试的几种 PA 信号中，包括来自 SV40、牛生长激素和最小兔 P-珠蛋白的信号，后者具有最高的转录活性（在 BeWo 和 JAR 细胞中分别是对照质粒的 3.9 倍和 26 倍）。添加第二个增强子增加了这些细胞中的转录活性。我们还发现可以通过选择表达盒来控制基因表达水平。这些结果对于 BeWo 和 JAR 细胞中的进一步转基因实验应该有用。 (c) 2006 Elsevier Ltd. 保留所有权利。
Nonviral gene delivery systems are useful for basic research in, trophoblasts. In these systems, gene expression is regulated by a cassette of regulatory elements within the plasmid, and the transcriptional activity differs among cell lines. In the present study, we used BeWo and JAR human trophoblast cell lines to systematically compare the transcriptional activities of several expression cassettes and those of a control plasmid made up of a simian virus 40 (SV40) promoter, a polyadenylation (PA) signal, and an enhancer. We also found that insertion of intron elements enhanced transcriptional activities in the following order: intron A > hybrid beta-globin-immunoglobin intron > no intron. Of several PA signals tested including those from SV40, bovine growth hormone, and the minimal rabbit P-globin, the latter had the highest transcriptional activities (3.9- and 26-fold over control plasmid in BeWo and JAR cells, respectively). Addition of a second enhancer increased the transcriptional activity in these cells. We also found that gene expression level can be controlled by selecting the expression cassette. These results should be useful for further transgene experiments in BeWo and JAR cells. (c) 2006 Elsevier Ltd. All rights reserved.