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使用mC-TAC-seq在碱基分辨率下分析RNA mC甲基化的实验方案。

Protocol for profiling RNA mC methylation at base resolution using mC-TAC-seq.

作者信息

Zhang Xiaoting, Lu Liang, Yi Chengqi, Li Xiaoyu

机构信息

State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China.

Department of Biochemistry and Department of Gastroenterology of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China; Institute of Immunology, Zhejiang University School of Medicine, Hangzhou 310058, China.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103599. doi: 10.1016/j.xpro.2025.103599. Epub 2025 Jan 31.

Abstract

RNA 5-methylcytosine (mC) is a widespread modification and plays a crucial role in gene expression regulation. Here, we present a protocol for transcriptome-wide mC methylome profiling at base resolution using bisulfite-free mC detection strategy enabled by ten-eleven translocation (TET)-assisted chemical labeling sequencing (mC-TAC-seq). We detail steps for TET-assisted chemical labeling, library construction, and data analysis. mC-TAC-seq enables accurate and robust mC detection in various RNA species. For complete details on the use and execution of this protocol, please refer to Lu et al..

摘要

RNA 5-甲基胞嘧啶(mC)是一种广泛存在的修饰,在基因表达调控中起着关键作用。在此,我们介绍一种使用由十-十一易位(TET)辅助化学标记测序(mC-TAC-seq)实现的无亚硫酸氢盐mC检测策略,在碱基分辨率下进行全转录组mC甲基化组分析的方案。我们详细说明了TET辅助化学标记、文库构建和数据分析的步骤。mC-TAC-seq能够在各种RNA物种中实现准确且可靠的mC检测。有关本方案使用和执行的完整详细信息,请参阅Lu等人的文献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4be6/11835647/14c9b03a217e/fx1.jpg

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