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对于RNA中的5-formylcytosine的定量测序方法

Ruitu Lyu¹, Kinga Pajdzik^1,2, Hui-Lung Sun¹

¹Department of Chemistry, The University of Chicago, Chicago, IL 60637, USA.

Israel journal of chemistry

|June 10, 2025

概括

研究人员开发了f5C-seq,这是一种在转录组中映射5-formylcytosine (f5C) 修饰的方法. 这项技术在RNA中发现了新的f5C位点,进步了我们对哺乳动物RNA修饰的理解.

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.
在RNA生物学,RNA生物学.

背景情况:

5-甲基细胞氨酸 (f5C) 是一种已知的RNA修饰,在特定的tRNA中发现.
哺乳动物中f5C的全转录组分布在很大程度上仍然没有表征.
现有的方法缺乏量化分辨率,无法在整个RNA景观中绘制f5C的地图.

研究的目的:

开发和验证一种新的测序方法,用于对5-formylcytosine (f5C) 的转录组全方位映射.
研究哺乳动物细胞中f5C修饰的分布和丰度.
确定新的f5C修饰RNA物种及其潜在的作用.

主要方法:

开发f5C-seq,一种定量测序技术,在逆转录过程中使用皮克降解和C-to-T突变签名.
使用已知的f5C位点在tRNA中验证f5C-seq,并评估ALKBH1-贫乏细胞中的修饰水平.
从HeLa和小鼠胚胎干细胞 (mESCs) 中应用f5C-seq来映射f5C在总RNA和染色质相关RNA (caRNA) 中.

主要成果:

f5C-seq在tRNA中成功识别和量化已知的f5C位点,在ALKBH1耗尽时显示显著减少.
该研究揭示了在HeLa和mESC两种线的转录组中全新的,高度修改的f5C位点.
适用于caRNA,在这个RNA分数中发现了以前未经表征的f5C修饰.

结论:

f5C-seq提供了一个强大的和定量平台,用于绘制哺乳动物全转录组范围的f5C修饰.
这些发现揭示了f5C在RNA中的分布比以前更广泛,包括在caRNA中.
这项工作为研究f5C在基因调节和细胞过程中的功能意义开辟了新的途径.

关键词:

5-甲基细胞氨酸的形式突变率的变化率是如何发生的量化测序是指量化的测序.阅读率通过率.转录组全方位的转录组.

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