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DpCoA标签Seq:通过马莱胺-醇反应进行直接纳米孔测序的dpCoA覆盖RNA条形码

Xiaojian Shao¹, Hailei Zhang², Zhou Zhu³

¹State Key Laboratory of Environmental and Biological Analysis, Department of Chemistry, Hong Kong Baptist University, Hong Kong, China.

Analytical chemistry

|July 13, 2023

概括

研究人员开发了dpCoA tagSeq来识别用3'-dephospho-coenzyme A (dpCoA) 封顶的RNA. 这种方法成功地在小鼠肝脏中识别了44个dpCoA封顶RNA基因,使得进一步的功能研究成为可能.

科学领域:

分子生物学分子生物学
在RNA生物学,RNA生物学.
生物化学生物化学

背景情况:

越来越多的人认可非正规的RNA封顶,包括尼古丁胺胺氨基二核酸 (NAD+) 和3'-dephospho-coenzyme A (dpCoA).
虽然已知dpCoA封闭的RNA,但这些dpCoA封闭的RNAs (dpCoA-RNAs) 的特定身份仍然难以捉摸.

研究的目的:

开发一种用于识别和表征dpCoA封顶RNA的新方法.
为了调查dpCoA-RNAs在哺乳动物系统中的患病率和身份.

主要方法:

开发了dpCoA tagSeq,一种采用醇反应性马莱胺基的方法,用条形码标记RNA标记dpCoA盖.
利用一个互补的DNA链来隔离标记的RNA,然后进行纳米孔测序.
在标签Seq.之前,通过将盖子降解为pyrophosphatase NudC来确认dpCoA的特异性.

主要成果:

dpCoA标签Seq有效地标记和捕获模型dpCoA-RNAs.
该方法证实了特异性,区分dpCoA盖和其他含有醇的分子.
在小鼠肝脏组织中鉴定了44个转录dpCoA-RNA的基因.

结论:

dpCoA tagSeq是一种有效的策略,用于识别和表征dpCoA-RNAs.
这种方法有助于未来对dpCoA caps的作用进行功能性调查.

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