iPSC 心脏分化过程中细胞周期基因 DNA 甲基化的动态可视化。

IF 3 4区医学 Q2 GENETICS & HEREDITY

Epigenomics Pub Date : 2024-12-01 Epub Date: 2024-11-27 DOI:10.1080/17501911.2024.2430171

Ning Li, Ba Thong Nguyen, Edward A Stitt, Zhenhe Zhang, W Robb MacLellan, Yiqiang Zhang

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引用次数: 0

摘要

背景：现有的传统分析方法对 DNA 甲基化的静态状态有深入的了解，但无法直观地显示表观遗传调控的动态变化：我们利用了一种基因组DNA甲基化报告基因（GMR）系统，该系统携带Snrpn最小启动子和Cdk1（细胞周期蛋白依赖性激酶1）或Sox2（SRY-Box转录因子2）的CpG区。小鼠 Sox2 GMR iPSC 在诱导心脏分化后迅速失去荧光报告信号。Cdk1 GMR报告信号在未分化的iPSC中很强，在心肌细胞分化过程中逐渐减弱。RT-qPCR和热序分析表明，在心脏分化过程中，Sox2和Cdk1的减少受其启动子CpG区超甲基化的调控：结论：GMR报告系统可用于以单细胞分辨率实时监测表观遗传学DNA修饰。

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Dynamic visualization of DNA methylation in cell cycle genes during iPSC cardiac differentiation.

Background: Existing analyses with conventional assays have generated significant insights into static states of DNA methylation but were unable to visualize the dynamics of epigenetic regulation.

Materials & results: We utilized a genomic DNA methylation reporter (GMR) system carrying Snrpn minimal promoter and CpG regions of Cdk1 (Cyclin-dependent kinase 1) or Sox2 (SRY-Box Transcription Factor 2). Mouse Sox2 GMR iPSCs rapidly lost fluorescent reporter signal upon the induction of cardiac differentiation. Cdk1 GMR reporter signal was strong in undifferentiated iPSCs, and gradually decreased during cardiomyocyte differentiation. RT-qPCR and pyrosequencing demonstrated that the reduction of Sox2 and Cdk1 was regulated by hypermethylation of their promoters' CpG regions during cardiac differentiation.

Conclusion: The GMR reporter system can be useful for monitoring real-time epigenetic DNA modification at single-cell resolution.

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来源期刊

Epigenomics GENETICS & HEREDITY-

CiteScore

5.80

自引率

2.60%

发文量

审稿时长

>12 weeks

期刊介绍： Epigenomics provides the forum to address the rapidly progressing research developments in this ever-expanding field; to report on the major challenges ahead and critical advances that are propelling the science forward. The journal delivers this information in concise, at-a-glance article formats – invaluable to a time constrained community. Substantial developments in our current knowledge and understanding of genomics and epigenetics are constantly being made, yet this field is still in its infancy. Epigenomics provides a critical overview of the latest and most significant advances as they unfold and explores their potential application in the clinical setting.