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mRNA Processing and Metabolism pp 223–243Cite as

Biochemical Dissection of RNA Silencing in Plants

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 257))

Abstract

Although RNA silencing was first discovered in plants, thus far it has been studied biochemically only in animals, where it is known as RNA interference (RNAi). In animals, two components of the RNAi pathway have been identified: Dicer, a multidomain RNase III that converts long double-stranded RNA (dsRNA) into small interfering RNA (siRNA) and the RNA-induced silencing complex (RISC), as siRNA-containing protein–RNA complex that targets complementary mRNA for destruction. We have developed methods for the biochemical dissection of plant RNA silencing. In this chapter, we describe in detail how to use wheat germ extract to study two distinct Dicer-like activities, RNA-dependent RNA polymerase (RdRP), and endogenous microRNA-programmed RISC activities. These comprehensive protocols should prove useful in the further dissection of the plant RNA silencing pathway, as well as for the validation of the predicted targets of endogenous plant microRNAs.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA

    Guiliang Tang & Phillip D. Zamore

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  1. Guiliang Tang
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  2. Phillip D. Zamore
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Editors and Affiliations

  1. Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH

    Daniel R. Schoenberg

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© 2004 Humana Press Inc., Totowa, NJ

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Tang, G., Zamore, P.D. (2004). Biochemical Dissection of RNA Silencing in Plants. In: Schoenberg, D.R. (eds) mRNA Processing and Metabolism. Methods in Molecular Biology™, vol 257. Humana Press. https://doi.org/10.1385/1-59259-750-5:223

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  • DOI: https://doi.org/10.1385/1-59259-750-5:223

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-225-4

  • Online ISBN: 978-1-59259-750-5

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