Targeted methylation of CMV and E1A viral promoters
Targeted methylation of CMV and E1A viral promoters
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DOI：
10.1016/j.bbrc.2010.09.131
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发表时间：
2010-11-12
影响因子：
3.1
通讯作者：
Hsiao, Shu-Huei
中科院分区：
文献类型：
--
作者：
Hsu, Chia-Chen;Li, Hsin-Pai;Hsiao, Shu-Huei
DNA甲基化是一种基因沉默和宿主防御系统，可以下调哺乳动物细胞中病毒基因的表达。用建立的靶向DNA甲基化方法证实了基因组整合的CMV和腺病毒5型EM启动子在体外被甲基化的病毒启动子片段导入MCF7和HEK293细胞后发生了高甲基化。在这两种情况下，靶向甲基化诱导的基因沉默可以通过添加5-氮杂-2‘-脱氧胞苷来逆转，证实CMV和E1A启动子受DNA甲基化调控。使用活细胞中的报告系统来确定靶向DNA甲基化的动力学。总之，靶向DNA甲基化能够有效地沉默敏感的病毒启动子，并为研究基因座特异性DNA甲基化对病毒基因表达的影响提供了一种替代策略。(C)2010 Elsevier Inc.保留所有权利。
DNA methylation is a gene-silencing and host defense system that can down-regulate viral gene expression in mammalian cells. An established targeted DNA methylation method was used to demonstrate that genome-integrated CMV and adenovirus type 5 EM promoters were hypermethylated after MCF7 and HEK293 cells were transfected with in vitro methylated viral promoter fragments. In both cases, the targeted methylation-induced gene silencing could be reversed by addition of 5-aza-2'-deoxycytidine, confirming that the CMV and E1A promoters are regulated by DNA methylation. The kinetics of the targeted DNA methylation was determined using a reporter system in live cells. In conclusion, targeted DNA methylation is able to efficiently silence susceptible viral promoters and provides an alternative strategy to study the impact of loci-specific DNA methylation in viral gene expression. (C) 2010 Elsevier Inc. All rights reserved.