College of Chemistry and Molecular Sciences, Key Laboratory of Biomedical Polymers-Ministry of Education, Wuhan University, Wuhan, Hubei 430072, PR China.
School of Public Health, Wuhan University, Wuhan, HuBei 430071, PR China.
Nucleic Acids Res. 2023 Sep 8;51(16):e87. doi: 10.1093/nar/gkad604.
Adenosine-to-inosine (A-to-I) RNA editing is a post-transcriptional processing event involved in diversifying the transcriptome and is responsible for various biological processes. In this context, we developed a new method based on the highly selective cleavage activity of Endonuclease V against Inosine and the universal activity of sodium periodate against all RNAs to enrich the inosine-containing RNA and accurately identify the editing sites. We validated the reliability of our method in human brain in both Alu and non-Alu elements. The conserved sites of A-to-I editing in human cells (HEK293T, HeLa, HepG2, K562 and MCF-7) primarily occurs in the 3'UTR of the RNA, which are highly correlated with RNA binding and protein binding. Analysis of the editing sites between the human brain and mouse brain revealed that the editing of exons is more conserved than that in other regions. This method was applied to three neurological diseases (Alzheimer's, epilepsy and ageing) of mouse brain, reflecting that A-to-I editing sites significantly decreased in neuronal activity genes.
腺嘌呤到次黄嘌呤（A-to-I）的 RNA 编辑是一种参与转录组多样化的转录后加工事件，负责各种生物过程。在这种情况下，我们开发了一种新方法，该方法基于内切酶 V 对次黄嘌呤的高度选择性切割活性和高碘酸钠对所有 RNA 的普遍活性，以富集含有次黄嘌呤的 RNA 并准确识别编辑位点。我们在人类大脑中的 Alu 和非 Alu 元件中验证了我们方法的可靠性。在人类细胞（HEK293T、HeLa、HepG2、K562 和 MCF-7）中，A-to-I 编辑的保守位点主要发生在 RNA 的 3'UTR 中，这与 RNA 结合和蛋白质结合高度相关。在人类大脑和老鼠大脑之间的编辑位点分析表明，外显子的编辑比其他区域更保守。该方法应用于三种神经疾病（阿尔茨海默病、癫痫和衰老）的老鼠大脑，反映出神经元活性基因中的 A-to-I 编辑位点显著减少。
