School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore, Singapore.
Genome Institute of Singapore, Agency for Science Technology and Research, Singapore, Singapore.
Methods Mol Biol. 2021;2181:163-176. doi: 10.1007/978-1-0716-0787-9_10.
Adenosine-to-inosine (A-to-I) RNA editing is a fundamental posttranscriptional mechanism that greatly diversifies the transcriptome in many living organisms, including mammals. Multiple studies have demonstrated the importance of this process not just in normal development and physiology but also in various human diseases. Importantly, the precise editing level of a site may have downstream consequences on cellular behavior. Hence, the editing levels should be quantified as accurately as possible. In this chapter, we describe how to examine RNA editing in human and mouse tissues. The rapid development of next-generation sequencing technologies is affording us an unprecedented ability to accurately measure the editing levels of numerous sites simultaneously. Our experimental workflow includes the harvesting of high-quality RNA samples and the construction of different high-throughput sequencing libraries. We also delineate the computational steps needed to analyze the sequencing data from an Illumina platform.
腺嘌呤到次黄嘌呤（A-to-I）RNA 编辑是一种基本的转录后机制，它极大地丰富了许多生物体的转录组，包括哺乳动物。多项研究表明，这一过程不仅在正常发育和生理过程中很重要，而且在各种人类疾病中也很重要。重要的是，一个位点的精确编辑水平可能会对细胞行为产生下游影响。因此，应该尽可能准确地量化编辑水平。在本章中，我们描述了如何检查人和小鼠组织中的 RNA 编辑。下一代测序技术的快速发展使我们能够前所未有地准确测量许多位点的编辑水平。我们的实验工作流程包括高质量 RNA 样本的采集和不同高通量测序文库的构建。我们还描述了从 Illumina 平台分析测序数据所需的计算步骤。
