使用纳米孔直接测序 RNA 和鉴定 RNA 修饰。
Direct Sequencing of RNA and RNA Modification Identification Using Nanopore.
机构信息
Division of Bioinformatics and Data Management for Research, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Department of Biomedical Informatics, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
出版信息
Methods Mol Biol. 2022;2477:71-77. doi: 10.1007/978-1-0716-2257-5_5.
Abstract
Direct RNA sequencing (dRNA-seq) simultaneously enables the detection of RNA modifications and characterization of full-length transcripts. In principle, full-length native RNA molecule is translocated through the nanopore by a motor protein while a sensor measures ionic current shifts. Then, the current shifts are interpreted by an algorithm that turn out to RNA sequence. Currently, the standard protocol of dRNA-seq provided by Oxford Nanopore Technologies (ONT) allows to directly ligate and sequence only polyadenylated RNA (poly(A) RNA). Here, we describe a method of dRNA-seq that can be applied for both poly(A) RNA and non-poly(A) tailed-RNA.
摘要
直接 RNA 测序（dRNA-seq）可同时检测 RNA 修饰和全长转录本的特征。从原理上讲，全长的天然 RNA 分子在通过纳米孔时被马达蛋白转运，同时传感器测量离子电流的变化。然后，算法将电流变化解释为 RNA 序列。目前，牛津纳米孔技术（ONT）提供的 dRNA-seq 标准方案仅允许直接连接和测序多聚腺苷酸化 RNA（poly(A) RNA）。在这里，我们描述了一种可应用于多聚腺苷酸化 RNA 和非多聚腺苷酸化 RNA 的 dRNA-seq 方法。
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