Quantitating plant microRNA-mediated target repression using a dual-luciferase transient expression system

Methods Mol Biol. 2015:1284:287-303. doi: 10.1007/978-1-4939-2444-8_14.

Abstract

microRNA (miRNA) mediated repression of target genes plays essential roles in a variety of functions in plants. An easy-to-use method that can effectively validate functional miRNA-target interactions in plants thus is of particular interest. Here, we describe an Agrobacterium tumefaciens-mediated in vivo assay utilizing a dual-luciferase reporter system. With this method, the strength of miRNA-mediated target repression can be quantified at both the mRNA (via qRT-PCR) and protein (via dual-luciferase assay) levels quickly and accurately.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens / genetics
  • Gene Expression Regulation, Plant
  • Gene Expression*
  • Genes, Reporter*
  • Luciferases / genetics*
  • MicroRNAs / genetics*
  • Plants / genetics*
  • RNA Interference*
  • RNA, Messenger / genetics*
  • Real-Time Polymerase Chain Reaction / methods

Substances

  • MicroRNAs
  • RNA, Messenger
  • Luciferases