Establishment of an In Vivo ARGONAUTE Reporter System in Plants

Methods Mol Biol. 2017:1640:73-91. doi: 10.1007/978-1-4939-7165-7_5.

Abstract

RNA silencing is not only an evolutionarily conserved gene regulatory mechanism, but in plants also serves as the basis for robust adaptive antiviral immune responses. ARGONAUTE (AGO) proteins form the catalytic cores of the RNA-guided ribonuclease complexes, which play a central role in RNA silencing. Here we describe an in vivo assay system for analyzing the activities of AGO proteins in the virological model plant Nicotiana benthamiana .

Keywords: ARGONAUTE; Agroinfiltratio n; Nicotiana benthamiana; RNA silencing; Transient reporter system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Agrobacterium / genetics*
  • Argonaute Proteins / analysis
  • Argonaute Proteins / genetics*
  • Blotting, Western / methods
  • Cloning, Molecular / methods*
  • DNA, Complementary / genetics
  • Electrophoresis, Polyacrylamide Gel / methods
  • Genetic Vectors / genetics
  • MicroRNAs / genetics
  • Models, Molecular
  • Nicotiana / genetics*
  • Plant Proteins / analysis
  • Plant Proteins / genetics*
  • RNA, Plant / genetics
  • Transformation, Genetic*

Substances

  • 3' Untranslated Regions
  • Argonaute Proteins
  • DNA, Complementary
  • MicroRNAs
  • Plant Proteins
  • RNA, Plant