Analysis of Methylation Status of Plant MicroRNAs

  • Susu Chen
  • Guodong RenEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1932)


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.

Key words

MicroRNA HEN1 Methylation β-elimination Denaturing PAGE Northern blot 



Work in the Ren laboratory is supported by grants from the National Key R&D Program of China (2016YFA0503200) and the National Natural Science Foundation of China (91740101, 31622009 and 31471221).

No conflict of interest was declared.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Genetic Engineering and Institute of Plant Biology, Department of Biochemistry, School of Life SciencesFudan UniversityShanghaiChina

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