Analysis of a plant transcriptional regulatory network using transient expression systems.

by Díaz-Triviño, Sara, Yuchen Long, Ben Scheres, Ikram Blilou
Year: 2017

Bibliography

Díaz-Triviño, Sara, Yuchen Long, Ben Scheres, and Ikram Blilou. "Analysis of a plant transcriptional regulatory network using transient expression systems." In Plant Gene Regulatory Networks, pp. 83-103. Humana Press, New York, NY, 2017.​​

Abstract

In plant biology, transient expression systems have become valuable approaches used routinely to rapidly study protein expression, subcellular localization, protein-protein interactions, and transcriptional activity prior to in vivo studies. When studying transcriptional regulation, luciferase reporter assays offer a sensitive readout for assaying promoter behavior in response to different regulators or environmental contexts and to confirm and assess the functional relevance of predicted binding sites in target promoters. This chapter aims to provide detailed methods for using luciferase reporter system as a rapid, efficient, and versatile assay to analyze transcriptional regulation of target genes by transcriptional regulators. We describe a series of optimized transient expression systems consisting of Arabidopsis thaliana protoplasts, infiltrated Nicotiana benthamiana leaves, and human HeLa cells to study the transcriptional regulations of two well-characterized transcriptional regulators SCARECROW (SCR) and SHORT-ROOT (SHR) on one of their targets, CYCLIN D6 (CYCD6).

Keywords

Promoter activity Transcriptional regulation Effectors Transfection dual luciferase reporter assay Luciferase