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Plant MicroRNAs pp 285–293Cite as

Analysis of Methylation Status of Plant MicroRNAs

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1932))

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.

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Acknowledgment

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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Authors and Affiliations

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

    Susu Chen & Guodong Ren

Authors
  1. Susu Chen
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  2. Guodong Ren
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Correspondence to Guodong Ren .

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Editors and Affiliations

  1. Unidad de Genómica Avanzada, Laboratorio Nacional de Genómica para la Biodiversidad (LANGEBIO), Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Irapuato, Guanajuato, Mexico

    Stefan de Folter

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Chen, S., Ren, G. (2019). Analysis of Methylation Status of Plant MicroRNAs. In: de Folter, S. (eds) Plant MicroRNAs. Methods in Molecular Biology, vol 1932. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9042-9_21

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  • DOI: https://doi.org/10.1007/978-1-4939-9042-9_21

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9041-2

  • Online ISBN: 978-1-4939-9042-9

  • eBook Packages: Springer Protocols

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