The tet-off system is more effective than the tet-on system for regulating transgene expression in a single adenovirus vector
The tet-off system is more effective than the tet-on system for regulating transgene expression in a single adenovirus vector
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DOI：
10.1002/jgm.261
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发表时间：
2002-05-01
影响因子：
3.5
通讯作者：
Hayakawa, T
中科院分区：
文献类型：
--
作者：
Mizuguchi, H;Hayakawa, T
背景在哺乳动物细胞中控制转基因表达对于基因治疗和基础研究中的基因功能研究是期望的。本研究评估了含有四环素可控表达系统的单腺病毒载体的功能（tet-off或tet-on系统）。方法产生含有tet-off或tet-on系统的Ad介导的二元转基因表达系统，其将具有四环素可调控启动子和四环素应答转录激活因子基因的目的基因引入E1和E3缺失区，分别在各种条件下，在体外和invivo.Results的腺病毒介导的tet-off和tet-on系统的功能进行了比较，包含tet-off系统的腺病毒载体提供了从20到500倍的基因表达的阴性对照，这取决于细胞类型和条件。与此相反，含有tet-on系统的Ad载体仅使基因表达增加2 ~ 28倍，并且需要高约2个对数数量级的诱导剂（强力霉素）来启动基因表达，与含有tet-off系统的Ad载体相比。结论含有tet-off系统的Ad载体比含有tet-on系统的Ad载体是更好的基因表达调控载体。含有四环素调控表达系统的单腺病毒载体将极大地促进基因功能的体外和体内分析，并可能用于基因治疗。版权所有（C）2002约翰威利父子有限公司
Background Control of transgene expression in mammalian cells is desirable for gene therapy and the study of gene function in basic research. This study evaluates the functionality of single adenovirus (Ad) vectors containing a tetracycline-controllable expression system (tet-off or tet-on system).Methods An Ad-mediated binary transgene expression system was generated containing a tet-off or a tet-on system, which introduced the gene of interest with a tetracycline-regulatable promoter and the tetracycline-responsive transcriptional activator gene into the E1 and E3 deletion regions, respectively. The functionality of the Ad-mediated tet-off and tet-on systems was compared under various conditions in vitro and in vivo.Results The Ad vector containing the tet-off system provided negative control of gene expression ranging from 20- to 500-fold, depending on the cell type and condition. In contrast, the Ad vector containing the tet-on system increased gene expression by only 2- to 28-fold and required about two-log orders higher concentration of an inducer (doxycycline) to switch on the gene expression, compared with the Ad vector containing the tet-off system.Conclusion Ad vectors containing the tet-off system are a better choice for regulated gene expression than Ad vectors containing the tet-on system. Single Ad vectors containing the tetracycline-controllable expression system will greatly facilitate in vitro and in vivo analyses of gene function and may be useful for gene therapy. Copyright (C) 2002 John Wiley Sons, Ltd.