Discovering A-to-I RNA Editing Through Chemical Methodology “ICE-seq”
Discovering A-to-I RNA Editing Through Chemical Methodology “ICE-seq”
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通过化学方法“ICE-seq”发现 A-to-I RNA 编辑
DOI：
10.1007/978-1-0716-0787-9_8
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发表时间：
2020
影响因子：
--
通讯作者：
Y.
中科院分区：
文献类型：
--
作者：
Sakurai;M.;Okada;S.;Ueda;H.;and Yang;Y.
RNA将腺苷编辑为肌苷，通过转录后调节基因表达，有助于广泛的生物学过程。为了理解这种效应，精确的肌苷映射是必要的。鉴定编辑位点最传统的方法是将cDNA序列与相应的基因组序列进行比较。然而，这种方法的错误发现率很高，因为鸟苷信号由于实验误差或获得的序列中的噪声而污染了被检测为鸟苷的真实肌苷信号。为了确保高准确性，我们开发了肌苷化学擦除（ICE）方法，利用肌苷氰乙化和逆转录- pcr准确生化鉴定RNA链中的肌苷。此外，我们将该技术应用于下一代测序技术，称为ICE-seq，对转录组中的A-to-I编辑位点进行无偏倚的全基因组筛选。
RNA editing of adenosines to inosines contributes to a wide range of biological processes by regulating gene expression post-transcriptionally. To understand the effect, accurate mapping of inosines is necessary. The most conventional method to identify an editing site is to compare the cDNA sequence with its corresponding genomic sequence. However, this method has a high false discovery rate because guanosine signals, due to experimental errors or noise in the obtained sequences, contaminate genuine inosine signals detected as guanosine. To ensure high accuracy, we developed the Inosine Chemical Erasing (ICE) method to accurately and biochemically identify inosines in RNA strands utilizing inosine cyanoethylation and reverse transcription-PCR. Furthermore, we applied this technique to next-generation sequencing technology, called ICE-seq, to conduct an unbiased genome-wide screening of A-to-I editing sites in the transcriptome.