Bidirectional promoter interference between two widely used internal heterologous promoters in a late-generation lentiviral construct
Bidirectional promoter interference between two widely used internal heterologous promoters in a late-generation lentiviral construct
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DOI：
10.1038/sj.gt.3303105
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发表时间：
2008-03-01
期刊：
影响因子：
5.1
通讯作者：
Ginn, S. L.
中科院分区：
文献类型：
--
作者：
Curtin, J. A.;Dane, A. P.;Ginn, S. L.
编码两个或多个基因的基因转移载体是潜在的强有力的研究工具，并准备在基因治疗应用中发挥越来越重要的作用。用于每个载体表达多于一个转基因的常见策略包括使用多个启动子、内部核糖体进入位点（IRES）元件、剪接信号和融合蛋白。其中，IRES元件和多启动子已被最广泛地使用。然而，多个启动子的使用可能受到启动子之间的干扰、启动子沉默和载体重排或缺失的影响。在这项研究中，我们证明了两个内部异源启动子之间的启动子干扰的情况下，晚一代慢病毒载体。干扰，涉及人巨细胞病毒立即早期启动子和人延伸因子-1 α启动子，发生双向的两个启动子显着损害相邻的转录单位的表达。所提供的数据不仅强调了这些广泛使用的启动子之间存在干扰的可能性，而且还强调了连续构建载体方法的价值，该方法可以识别此类效应。
Gene transfer vectors encoding two or more genes are potentially powerful research tools and are poised to play an increasingly important role in gene therapy applications. Common strategies employed to express more than one transgene per vector include the use of multiple promoters, internal ribosome entry site (IRES) elements, splicing signals and fusion proteins. Of these, the IRES elements and multiple promoters have been most widely used. The use of multiple promoters, however, may be compromised by interference between promoters, promoter silencing and vector rearrangements or deletions. In this study, we demonstrate promoter interference between two internal heterologous promoters in the context of a late-generation lentiviral vector. The interference, involving the human cytomegalovirus-immediate-early promoter and human elongation-factor-1 alpha promoter, occurred bidirectionally with both promoters markedly impairing expression of the adjacent transcription unit. The data presented not only highlight the potential for interference between these widely-used promoters, but also the value of a sequential approach to vector construction that allows such effects to be recognized.