Adenosine-to-inosine (A-to-I) RNA editing is a widespread and conserved post-transcriptional modification, producing significant changes in cellular function and behavior. Accurately identifying, detecting, and quantifying these sites in the transcriptome is necessary to improve our understanding of editing dynamics, its broader biological roles, and connections with diseases. Chemical labeling of edited bases coupled with affinity enrichment has enabled improved characterization of several forms of RNA editing. However, there are no approaches currently available for pull-down of inosines. To address this need, we explore acrylamide as a labeling motif and report here an acrylamidofluorescein reagent that reacts with inosine and enables enrichment of inosine-containing RNA transcripts. This method provides improved sensitivity in the detection and identification of inosines toward a more comprehensive transcriptome-wide analysis of A-to-I editing. Acrylamide derivatization is also highly generalizable, providing potential for the labeling of inosine with a wide variety of probes and affinity handles.

中文翻译：

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化学标记和使用丙烯酰胺基荧光素亲和捕获肌苷的RNA。

腺苷到肌苷（A到I）RNA编辑是一种广泛且保守的转录后修饰，在细胞功能和行为方面产生了显着变化。准确地识别，检测和量化转录组中的这些位点对于提高我们对编辑动态，其更广泛的生物学作用以及与疾病的联系的理解是必要的。结合亲和力富集的化学修饰碱基的化学标记已使几种形式的RNA编辑的表征得以改善。但是，目前没有可用于下拉肌苷的方法。为了满足这一需求，我们探索了丙烯酰胺作为标记基序的方法，并在此报告了一种与肌苷反应并能够富集肌苷的RNA转录物的丙烯酰胺荧光素试剂。此方法可提高肌苷检测和鉴定的灵敏度，从而实现A-to-I编辑的更全面的转录组范围分析。丙烯酰胺衍生化也具有很高的普遍性，为使用各种探针和亲和柄标记肌苷提供了潜力。