MePMe-seq:通过代谢propargyl标记和测序在mRNA中绘制无抗体同时m6A和m5C的映射
Katja Hartstock1, Nadine A Kueck1, Petr Spacek1
1Institute of Biochemistry, Faculty of Chemistry and Pharmacy, University of Münster, Corrensstraße 36, 48149, Münster, Germany.
Nature communications
|November 7, 2023
概括
研究人员开发了一种新方法,使用可点击的SAM前体来检测和绘制mRNA甲基化位点,包括N6-甲基氨酸 (m6A) 和5-甲基氨酸 (m5C),以单核酸精度.
科学领域:
- 分子生物学分子生物学
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
- 生物化学 生物化学
背景情况:
- mRNA的内部修改,主要是甲基化,在转录后调节基因表达.
- S-adenosyl-L-methionine (SAM) 是这些甲基化过程的关键代谢枢纽.
研究的目的:
- 开发一种用于检测和识别各种mRNA甲基化位点的新方法.
- 为了使用下一代测序,精确地绘制甲基化位点.
主要方法:
- 使用可点击的SAM前体propargyl-selenohomocysteine (PSH) 的代谢标签.
- 一个SAM模拟的细胞内生成导致propargylated核化物.
- 与MePMe-seq的下一代测序集成,以映射N6-甲基氨酸 (m6A) 和5-甲基胺 (m5C) 位点.
主要成果:
- 成功检测和识别各种mRNA甲基化位点.
- 在mRNA中精确地绘制m6A和m5C位点的单核酸精度映射.
- 使用终端配置分析区分m6A与A_m和m1A位点.
结论:
- MePMe-seq克服了以前基于抗体和序列动机的方法的局限性.
- 可点击的基于SAM的代谢标记可促进跨RNA,DNA和蛋白质的甲基化联合评估.
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