The TARGET System: Rapid Identification of Direct Targets of Transcription Factors by Gene Regulation in Plant Cells

Matthew D. Brooks, Kelsey M. Reed, Gabriel Krouk, Gloria M. Coruzzi, Bastiaan O.R. Bargmann

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The TARGET system allows for the rapid identification of direct regulated gene targets of transcription factors (TFs). It employs the transient transformation of plant protoplasts with inducible nuclear entry of the TF and subsequent transcriptomic and/or ChIP-seq analysis. The ability to separate direct TF–target gene regulatory interactions from indirect downstream responses and the significantly shorter amount of time required to perform the assay, compared to the generation of transgenics, make this plant cell-based approach a valuable tool for a higher throughput approach to identify the genome-wide targets of multiple TFs, to build validated transcriptional networks in plants. Here, we describe the use of the TARGET system in Arabidopsis seedling root protoplasts to map the gene regulatory network downstream of transcription factors-of-interest.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages1-12
Number of pages12
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2594
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Fluorescence activated cell sorting
  • Gene regulatory network
  • Protoplast
  • RNA-seq
  • Transcription factor

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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