Functional Analysis of Promoter CpG Methylation Using a CpG-Free Luciferase Reporter Vector
Functional Analysis of Promoter CpG Methylation Using a CpG-Free Luciferase Reporter Vector
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DOI：
10.4161/epi.1.3.3327
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发表时间：
2006-07-01
期刊：
影响因子：
3.7
通讯作者：
Rehli, Michael
中科院分区：
文献类型：
--
作者：
Klug, Maja;Rehli, Michael
近端启动子内CpG二核苷酸的甲基化通常与转录沉默相关。高度甲基化的CpG岛启动子的甲基化依赖性阻遏已被充分确立，其特征在于高密度的CpG残基。CpG DNA甲基化对CpG-贫启动子的影响不太清楚，可能是由于缺乏方便的体外检测启动子活性的测定系统。在这份报告中，我们描述了一种新的荧光素酶报告载体，pCpGL，它完全缺乏CpG二核苷酸，可用于研究在转染试验中启动子DNA甲基化的影响。而传统的报告载体，含有大量的骨干CpG残基显着抑制CpG-无启动子甲基化时，我们的新的报告载体仅被抑制，由于功能上重要的，甲基化的CpG残基的存在。pCpGL载体为研究CpG甲基化对富CpG和贫CpG启动子的影响提供了有用的工具。
Methylation of CpG dinucleotides within proximal promoters is often associated with transcriptional silencing. Methylation-dependent repression is well established for hypermethylated CpG island promoters that are characterized by a high density of CpG residues. The effect of CpG DNA methylation on CpG-poor promoters is less well characterized, probably due to the lack of convenient assay systems to test promoter activities in vitro. In this report, we describe a novel luciferase reporter vector, pCpGL, which completely lacks CpG dinucleotides and can be used to study the effect of promoter DNA methylation in transfection assays. Whereas a traditional reporter vector that contains a large number of backbone CpG residues significantly represses a CpG-free promoter when methylated, our new reporter vector is only repressed due to the presence of functionally important, methylated CpG residues. The pCpGL vector provides a useful tool to study the effects of CpG methylation on CpG-rich and CpG-poor promoters.