Development of an in vivo assay system for aberrant DNA methylation

异常 DNA 甲基化体内检测系统的开发

• 批准号: 23650589
• 负责人: TAKADA ERIKO
• 金额: $ 2.41万
• 依托单位: National Center of Neurology and Psychiatry
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23650589/
• 关键词: エピジェネティクス; DNAメチル化; 培養細胞; LacI; 検出系; 検出; トランスジェニックマウス

Our knowledge of the factors that induce aberrant DNA methylation is limited. The present study aimed to develop an assay system to detect aberrant DNA methylation in vivo. The vector, in which LacI was inserted into downstream of the promoter CpG island, and the vector, in which EGFP as the reporter gene was inserted into downstream of lacO, were stably introduced into a human colon cancer cell line. The cells induced aberrant methylation of the promoter with green fluorescence during xenograft tumor formation in nude mice. In the tumor without fluorescence, no methylation was induced. These results indicate that cell lines constructed in this study may have the ability to detect induction of aberrant methylation. In the next step, construction of transgenic mouse using this strategy would be expected.

我们对诱导异常DNA甲基化的因素的了解是有限的。 本研究旨在建立一种检测体内DNA甲基化的方法。 将LacI插入启动子CpG岛下游的载体和作为报告基因的EGFP插入lacO下游的载体稳定导入人结肠癌细胞系。 在裸鼠异种移植瘤形成过程中，这些细胞诱导了具有绿色荧光的启动子的异常甲基化。 在没有荧光的肿瘤中，没有诱导甲基化。 这些结果表明，本研究中构建的细胞系可能具有检测异常甲基化诱导的能力。 在下一步的工作中，利用该策略构建转基因小鼠是值得期待的。