Analysis of Methylation Status of Plant MicroRNAs

Susu Chen; Guodong Ren

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

Analysis of Methylation Status of Plant MicroRNAs

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

Authors

Susu Chen¹, Guodong Ren²

Affiliations

¹ State Key Laboratory of Genetic Engineering and Institute of Plant Biology, Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai, China.
² State Key Laboratory of Genetic Engineering and Institute of Plant Biology, Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai, China. gdren@fudan.edu.cn.

PMID: 30701508
DOI: 10.1007/978-1-4939-9042-9_21

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.

Keywords: Denaturing PAGE; HEN1; Methylation; MicroRNA; Northern blot; β-elimination.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / genetics
Arabidopsis Proteins / genetics
Methylation
Methyltransferases / genetics*
MicroRNAs / genetics*
RNA Stability / genetics
RNA, Plant / genetics*
RNA, Small Interfering / genetics
RNA, Small Untranslated / genetics

Substances

Arabidopsis Proteins
MicroRNAs
RNA, Plant
RNA, Small Interfering
RNA, Small Untranslated
Methyltransferases