核m6A-Label-Seq使转录组范围内的核m6A在单基分辨率下进行分析
Chenyang Huang1, Xiner Ying1, Xiao Shu2
1MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310058, China.
ACS chemical biology
|March 10, 2026
概括
核m6A-label-seq能够精确地映射核非编码RNA中的N6M-甲基氨酸 (m6A). 这种方法提供了核m6的高分辨率分析,具有减少输入和更快的图书馆构建的表转录组.
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:
- 在mRNA调节中,N6 - - 甲基氨酸 (m6A) 修饰至关重要,但它在核非编码RNA中的作用不太了解.
- 现有的m6A分析方法缺乏单基分辨率,或者没有针对核RNA进行优化.
研究的目的:
- 开发和验证一种新的方法,Nuclear-m6A-label-seq,用于核非编码RNA中m6A的高分辨率,全转录组映射.
- 为高效的核m6的详细协议提供A表表转录组概况.
主要方法:
- 核-m6A-label-seq利用在腺位上用基组进行代谢标记,然后转化为循环腺 (cyc-A).
- 艾滋病毒逆转录酶在cDNA合成过程中诱导循环-A位点的基因错误整合,使单基因分辨率映射成为可能.
- 该协议包括连续核RNA隔离,rRNA枯竭和使用rMATS-DVR管道进行分析.
主要成果:
- 与以前的方法相比,核-m6A-label-seq显著减少了RNA输入 (5μg) 和库建设时间 (~6h).
- 该方法捕获了多基和非多基核转录.
- 通过将读数与T2T-CHM13人类基因组对齐,可以准确地绘制m6A位点,包括重复区域.
结论:
- 核m6A-label-seq是一种直接的,高分辨率的,高效的方法,用于对核m6A表转录组进行分析.
- 这种方法有助于研究核非编码RNA中的m6A功能.
- 详细的协议使得表表表转录学研究能够得到更广泛的采用和推进.
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