The impact of YTHDF2 knockout on the distribution of 5-methylcytosine in RNA

5-methylcytosine is an important epigenetic modification found in both DNA and RNA. Transcriptome-wide mapping of 5-methylcytosine revealed that, after CRISPR-Cas9-mediated knockout of YTHDF2 gene, 545 sites displayed significant changes in 5-methylcytosine levels, with the majority of 5-methylcytosine sites in ribosomal RNA exhibiting substantially increased levels of methylation. Our data demonstrateda broader role of YTHDF2 in the post-transcriptional regulation of RNA. Overall design: Combine RNA bisulfite conversion with next-generation sequencing to monitor the variation of 5-methylcytosine in the transcriptome of HEK293T and isogenic cells with YTHDF2 gene being knocked out.

5-甲基胞嘧啶（5-methylcytosine）是广泛存在于DNA与RNA中的重要表观遗传修饰。对5-甲基胞嘧啶开展转录组范围的定位分析显示，经CRISPR-Cas9介导的YTHDF2基因敲除后，共有545个位点的5-甲基胞嘧啶水平出现显著变化，其中核糖体RNA（ribosomal RNA）中的绝大多数5-甲基胞嘧啶位点的甲基化水平均显著升高。本研究数据表明，YTHDF2在RNA的转录后调控中发挥着更为广泛的作用。实验设计概述：将RNA亚硫酸氢盐转化（RNA bisulfite conversion）技术与下一代测序（next-generation sequencing）相结合，用于监测HEK293T细胞及其YTHDF2基因敲除的同基因细胞转录组中5-甲基胞嘧啶的水平变化。