CRES-T, an effective gene silencing system utilizing chimeric repressors

Methods Mol Biol. 2011:754:87-105. doi: 10.1007/978-1-61779-154-3_5.

Abstract

Chimeric REpressor gene Silencing Technology (CRES-T) is a useful tool for functional analysis of plant transcription factors. In this system, a chimeric repressor that is produced by fusion of a transcription factor to the plant-specific EAR-motif repression domain (SRDX) suppresses target genes of a transcription factor dominantly over the activity of endogenous and functionally redundant transcription factors. As a result, the transgenic plants that express a chimeric repressor exhibit phenotypes similar to loss-of-function of the alleles of the gene encoding the transcription factor. This system is simple and effective and can be used as a powerful tool not only for functional analysis of redundant transcription factors but also for the manipulation of plant traits by active suppression of the gene expression. Strategies for construction of the chimeric repressors and their expression in transgenic plants are described. Transient effector-reporter assays for functional analysis of transcription factors and detection of protein-protein interactions using the trans-repressive activity of SRDX repression domain are also described.

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism
  • Gene Expression
  • Gene Silencing*
  • Mutant Chimeric Proteins / genetics*
  • Mutant Chimeric Proteins / metabolism
  • Promoter Regions, Genetic
  • Repressor Proteins / genetics*
  • Repressor Proteins / metabolism
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism*

Substances

  • Arabidopsis Proteins
  • Mutant Chimeric Proteins
  • Repressor Proteins
  • Transcription Factors