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使用纳米-DMS-MaP确定异形特异性RNA结构

Anne-Sophie Gribling-Burrer¹, Patrick Bohn², Redmond P Smyth^3,4

¹Helmholtz Institute for RNA-based Infection Research, Helmholtz Centre for Infection Research, Würzburg, Germany. anne-sophie.gribling@helmholtz-hiri.de.

Nature protocols

|February 12, 2024

概括

纳米孔二甲基硫酸盐突变概况 (Nano-DMS-MaP) 确定特定异构体的RNA结构. 这种方法使用纳米孔测序揭示了长,类似的RNA分子中隐藏的结构差异.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
生物化学生物化学

背景情况:

了解RNA结构对于阐明其生物功能至关重要.
细胞表达多种RNA异型,其序列因替代拼接和差异多化等过程而变化.
现有的RNA结构确定方法经常与长且高度相似的RNA分子相斗争.

研究的目的:

描述纳米孔二甲基硫酸盐突变分析 (Nano-DMS-MaP) 的开发和应用.
为了在现场进行异构体特异性RNA结构的确定.
解决长和高度相似的RNA分子中的结构差异.

主要方法:

使用纳米孔测序进行长读RNA结构分析.
使用二甲基硫酸盐突变分析 (DMS-MaP) 来进行RNA结构的化学探测.
提供了一个从实验设计和执行到数据分析的协议.

主要成果:

在复杂的混合物中,纳米-DMS-MaP成功地解决了单个异构体的RNA结构.
该方法揭示了高度相似的RNA分子中以前隐藏的结构变异.
对于经验丰富的研究人员来说,细胞内探测实验在3-4天内是可行的.

结论:

纳米-DMS-MaP是一种强大的工具,用于确定异形特异性RNA结构.
这种方法克服了用于分析复杂RNA结构的短读序列的局限性.
该协议促进了纳米-DMS-MaP的应用,用于全面的RNA结构研究.

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