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Antisense probing of dynamic RNA structures
Methods ( IF 4.8 ) Pub Date : 2020-11-01 , DOI: 10.1016/j.ymeth.2020.01.015
Alexandra J Lukasiewicz 1 , Lydia M Contreras 2
Affiliation  

RNA regulation is influenced by the dynamic changes in conformational accessibility on the transcript. Here we discuss the initial validation of a cell-free antisense probing method for structured RNAs, using the Tetrahymena group I intron as a control target. We observe changes in signal that qualitatively match prior traditional DMS footprinting experiments. Importantly, we have shown that application of this technique can elucidate new RNA information given its sensitivity for detecting rare intermediates that are not as readily observed by single-hit kinetics chemical probing techniques while elucidating new information about the RNA conformation. Observing changes in RNA accessibility has broad applications in determining the effect that regulatory elements have on regional structures. We speculate that this method could be useful in quickly observing those interactions, along with other phenomena that influence RNA accessibility including RNA-RNA interactions and small molecules.

中文翻译:

动态RNA结构的反义探测

RNA 调控受转录本构象可及性的动态变化影响。在这里,我们讨论了对结构化 RNA 的无细胞反义探测方法的初步验证,使用四膜虫 I 组内含子作为对照目标。我们观察到与先前传统 DMS 足迹实验定性匹配的信号变化。重要的是,我们已经表明,该技术的应用可以阐明新的 RNA 信息,因为它具有检测稀有中间体的敏感性,这些中间体在阐明有关 RNA 构象的新信息时不容易通过单次动力学化学探测技术观察到。观察 RNA 可及性的变化在确定监管要素对区域结构的影响方面具有广泛的应用。
更新日期:2020-11-01
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