Chen Susu, Ren Guodong
State Key Laboratory of Genetic Engineering and Institute of Plant Biology, Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai, China.
Methods Mol Biol. 2019;1932:285-293. doi: 10.1007/978-1-4939-9042-9_21.
Small noncoding RNAs of 20-30 nucleotides in length are key mediators of gene silencing. 2'-O-Methylation on the 3' terminal nucleotide of several types of small RNAs, including microRNAs (miRNAs) and small interfering RNAs (siRNAs) in plants, PIWI-interacting RNAs (piRNAs) in animals, and some siRNAs in Drosophila and Caenorhabditis elegans, provides a key protective mechanism against 3' tailing- and trimming-mediated destabilization. The methylation reaction is catalyzed by the small RNA methyltransferase HUA ENHANCER 1 (HEN1). In this chapter, we describe a detailed protocol for analyzing 3' end methylation status of plant miRNAs, which can be applicable to other types of small RNAs as well.
长度为20 - 30个核苷酸的小非编码RNA是基因沉默的关键介质。几种类型的小RNA，包括植物中的微小RNA（miRNA）和小干扰RNA（siRNA）、动物中的PIWI相互作用RNA（piRNA）以及果蝇和秀丽隐杆线虫中的一些siRNA，其3'末端核苷酸上的2'-O-甲基化提供了一种关键的保护机制，以防止3'尾巴化和修剪介导的去稳定化。甲基化反应由小RNA甲基转移酶华增强子1（HEN1）催化。在本章中，我们描述了一种详细的方案，用于分析植物miRNA的3'末端甲基化状态，该方案也可适用于其他类型的小RNA。
