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In Situ Localization of Small RNAs in Plants

  • Cristina F. Marco
  • Damianos S. Skopelitis
  • Marja C. P. TimmermansEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1932)

Abstract

Small RNAs have vital roles in numerous aspects of plant biology. Deciphering their precise contributions requires knowledge of a small RNA’s spatiotemporal pattern of accumulation. The in situ hybridization protocol described here takes advantage of locked nucleic acid (LNA) oligonucleotide probes to visualize small RNA expression at the cellular level with high sensitivity and specificity. The procedure is optimized for paraffin-embedded plant tissue sections, is applicable to a wide range of plants and tissues, and can be completed within 2–6 days.

Key words

Small RNA microRNA In situ hybridization Tissue specificity Plant development Cell-to-cell communication 

Notes

Acknowledgments

Damianos Skopelitis was supported by an HFSP long-term postdoctoral fellowship (LT000257/2009). Work on small RNA regulation in the Timmermans lab is supported by grants from the National Science Foundation (IOS-1355018), the Deutsche Forschungsgemeinschaft (SFB 1101 project C06), and an Alexander von Humboldt Professorship.

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Copyright information

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

Authors and Affiliations

  • Cristina F. Marco
    • 1
  • Damianos S. Skopelitis
    • 1
  • Marja C. P. Timmermans
    • 1
    • 2
    Email author
  1. 1.Cold Spring Harbor LaboratoryCold Spring HarborUSA
  2. 2.Center for Plant Molecular BiologyUniversity of TübingenTübingenGermany

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