诱导多能干细胞向心肌细胞分化过程中细胞周期基因DNA甲基化的动态可视化
Dynamic visualization of DNA methylation in cell cycle genes during iPSC cardiac differentiation.
作者信息
Li Ning, Nguyen Ba Thong, Stitt Edward A, Zhang Zhenhe, MacLellan W Robb, Zhang Yiqiang
机构信息
Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA, USA.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
出版信息
Epigenomics. 2024 Dec-Dec;16(23-24):1407-1414. doi: 10.1080/17501911.2024.2430171. Epub 2024 Nov 27.
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 (Cyclin-dependent kinase 1) or (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 and 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.
背景
现有的传统检测分析对DNA甲基化的静态状态有了重要见解，但无法可视化表观遗传调控的动态过程。
材料与结果
我们利用了一个基因组DNA甲基化报告基因（GMR）系统，该系统携带小核核糖核蛋白（Snrpn）最小启动子以及细胞周期蛋白依赖性激酶1（Cdk1）或SRY盒转录因子2（Sox2）的CpG区域。小鼠Sox2 GMR诱导多能干细胞（iPSC）在诱导心脏分化时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，并在心肌细胞分化过程中逐渐降低。逆转录定量聚合酶链反应（RT-qPCR）和焦磷酸测序表明，在心脏分化过程中，Cdk1和Sox2的减少是由其启动子CpG区域的高甲基化调节的。
结论
GMR报告系统可用于在单细胞分辨率下监测实时表观遗传DNA修饰。
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