系统地描述了与身体相关的mRNAs的组成和处理动态
Zhiyuan Sun1,2,3,4, Xiaozhen Wen1,2,3,4, Yanping Li1,2,3,4
1Shenzhen Key Laboratory of Gene Regulation and Systems Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, China.
Nature communications
|November 10, 2025
概括
我们开发了PB-TRIBE-STAMP,用于在处理器官 (PB) 中分析RNA. 这种方法确定了成千上万的PB相关的mRNA,揭示了RNA调节和这些细胞结构内的动态.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 在RNA生物学,RNA生物学.
背景情况:
- 处理体 (PBs) 是关键的动态,无膜有机体,参与RNA调节.
- 现有的与PBs相关的RNA分析方法有限,阻碍了全面的分析.
- 了解PB相关RNA是解读转录后基因调节的关键.
研究的目的:
- 开发和验证一种用于加工机构内的RNA系统分析的新方法.
- 在人类细胞系中识别和描述与PBs相关的信使RNA (mRNA).
- 研究PB关联RNA的特征和动态,包括多分子A尾长和3' UTR异型的使用.
主要方法:
- 开发PB-TRIBE-STAMP,一种使用两个正交RNA编辑酶 (APOBEC1-DDX6和LSM14A-ADAR2dd) 进行RNA分析的工具.
- 同时在HCT116和HEK293T细胞中应用PB-TRIBE-STAMP,以识别PB相关的mRNA.
- 生物化学分离PBs,然后进行RNA测序 (RNA-seq) 进行验证.
- 与长读序列的集成,以分析多类A尾长度.
- 建立TRIBE-ID和单细胞LSM14A-TRIBE-ID (sc-LSM14A-TRIBE-ID) 用于高分辨率的时间和单细胞分析.
主要成果:
- 在HCT116和HEK293T细胞中,PB-TRIBE-STAMP分别发现了1,639个和2,577个PB相关的mRNA.
- 验证证实了在孤立的PB中编辑成绩单的丰富性.
- 发现与PB相关的转录具有较短的多A尾,其中约有3' UTR异型表现出特定的PB关联.
- 在未折叠蛋白质反应 (UPR) 期间,TRIBE-ID揭示了LSM14A相关的XBP1转录的更高拼接效率.
- sc-LSM14A-TRIBE-ID在细胞周期进展过程中展示了动态mRNA-LSM14A/PB关联模式.
结论:
- PB-TRIBE-STAMP是一种有效的工具,用于对PB相关RNA进行全面的分析.
- 这项研究揭示了PB相关的mRNA的新特性,包括多A尾长和3' UTR异型特异性.
- mRNA-LSM14A/PB关联的动态调节发生在细胞应激 (UPR) 和细胞周期进展期间.
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