植物微小RNA甲基化状态分析

Analysis of Methylation Status of Plant MicroRNAs.

作者信息

Chen Susu, Ren Guodong

机构信息

State Key Laboratory of Genetic Engineering and Institute of Plant Biology, Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai, China.

出版信息

Methods Mol Biol. 2019;1932:285-293. doi: 10.1007/978-1-4939-9042-9_21.

DOI:10.1007/978-1-4939-9042-9_21

PMID:30701508

Abstract

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.

摘要

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。