In Situ Localization of Small RNAs in Plants

Methods Mol Biol. 2019:1932:159-173. doi: 10.1007/978-1-4939-9042-9_12.

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.

Keywords: Cell-to-cell communication; In situ hybridization; Plant development; Small RNA; Tissue specificity; microRNA.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • In Situ Hybridization / methods
  • MicroRNAs / genetics*
  • Oligonucleotide Probes / genetics
  • Oligonucleotides / genetics
  • Plants / genetics*
  • RNA, Plant / genetics*
  • Sensitivity and Specificity

Substances

  • MicroRNAs
  • Oligonucleotide Probes
  • Oligonucleotides
  • RNA, Plant
  • locked nucleic acid