Training Program in Neuroscience, Vanderbilt University School of Medicine, Nashville, TN, USA.
Department of Molecular Physiology & Biophysics, Vanderbilt University School of Medicine, Nashville, TN, USA.
Methods Mol Biol. 2021;2181:97-111. doi: 10.1007/978-1-0716-0787-9_7.
The conversion of adenosine to inosine (A to I) by RNA editing represents a common posttranscriptional mechanism for diversification of both the transcriptome and proteome, and is a part of the cellular response for innate immune tolerance. Due to its preferential base-pairing with cytosine (C), inosine (I) is recognized as guanosine (G) by reverse transcriptase, as well as the cellular splicing and translation machinery. A-to-I editing events appear as A-G discrepancies between genomic DNA and cDNA sequences. Molecular analyses of RNA editing have leveraged these nucleoside differences to quantify RNA editing in ensemble populations of RNA transcripts and within individual cDNAs using high-throughput sequencing or Sanger sequencing-derived analysis of electropherogram peak heights. Here, we briefly review and compare these methods of RNA editing quantification, as well as provide experimental protocols by which such analyses may be achieved.
通过 RNA 编辑将腺苷转化为肌苷（A 到 I）是转录组和蛋白质组多样化的常见转录后机制，也是细胞先天免疫耐受反应的一部分。由于肌苷（I）与胞嘧啶（C）的优先碱基配对，它被逆转录酶以及细胞剪接和翻译机制识别为鸟嘌呤（G）。A 到 I 编辑事件在基因组 DNA 和 cDNA 序列之间表现为 A-G 差异。RNA 编辑的分子分析利用这些核苷差异，使用高通量测序或基于电泳图谱峰高的 Sanger 测序衍生分析，在 RNA 转录物的整体群体和单个 cDNA 中定量 RNA 编辑。在这里，我们简要回顾和比较了这些 RNA 编辑定量方法，并提供了可以实现这些分析的实验方案。
