Construction and validation of a novel dual reporter vector for studying mammalian bidirectional promoters
Construction and validation of a novel dual reporter vector for studying mammalian bidirectional promoters
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DOI：
10.1016/j.plasmid.2014.05.001
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发表时间：
2014-07-01
期刊：
影响因子：
2.6
通讯作者：
Rocha, Dominique
中科院分区：
文献类型：
--
作者：
Lejard, Veronique;Rebours, Emmanuelle;Rocha, Dominique
基因表达调控在细胞功能中起着重要作用。随着测序技术的发展，越来越多的基因组可用，对可能影响基因表达调控的基因组结构进行全基因组分析成为可能。对几个基因组的分析发现了一类调控区域，其中包含启动两个不同基因转录的元件，这些基因在两个相反的方向上以头对头的方式排列。这些调控区域被称为双向启动子。虽然双向启动子已被发现多年，但最近的基因组规模研究表明，高达10%的基因表达的调控是由双向启动子控制的。这些发现主要基于计算工作，只有有限数量的假定双向启动子得到了实验验证。发展研究双向启动子的方法将使研究人员了解这些区域是如何被调节的，以及发散转录在基因表达中所起的作用。在此，我们开发了一种新的双荧光报告基因载体来研究哺乳动物双向启动子的转录输出。我们证明该载体能够在双向启动子的控制下表达报告基因，使用已知的人类OSGEP/APEX双向启动子。(C) 2014爱思唯尔公司版权所有。
Regulation of gene expression plays important role in cellular functions. With the development of sequencing techniques, more and more genomes are available and genome-wide analyses of genomic structures that may affect gene expression regulation are now possible. Analyses of several genomes have found a class of regulatory regions that contain elements that initiate transcription of two different genes positioned with a head-to-head arrangement in two opposite directions. These regulatory regions are known as bidirectional promoters. Although bidirectional promoters have been known for years, recent genome-scale studies have shown that the regulation of the expression of up to 10% of the genes are controlled by bidirectional promoters. These findings are based mostly on computational work and only a limited number of putative bidirectional promoters have been experimentally validated. Developing methods to study bidirectional promoters will allow researchers to understand how these regions are regulated and the roles that divergent transcription plays in the expression of genes. Here, we have developed a novel dual-fluorescence reporter gene vector to study the transcriptional output of mammalian bidirectional promoters. We demonstrate that this vector is capable of expressing reporter genes under the control of bidirectional promoters, using the known human OSGEP/APEX bidirectional promoter. (C) 2014 Elsevier Inc. All rights reserved.