title: High-Throughput Protoplast Trans-Activation (PTA) Screening to Define Transcription Factors in Auxin-Mediated Gene Regulation.
abstract: Genome sequencing and annotation studies clearly highlight the impact of transcriptional regulation in plants. However, functional characterization of the majority of transcriptional regulators remains elusive. Hence, high-throughput techniques are required to facilitate their molecular analysis. Here, we provide a detailed protocol to conduct a high-throughput protoplast trans-activation (PTA) screening, which enables simultaneous analysis of up to 95 individual transcription factor activities on a customizable promoter:LUCIFERASE reporter. This system is well suited to decipher complex transcriptional networks such as that triggered by the phytohormone auxin.
authors.affiliation: Pharmazeutische Biologie, Julius-von-Sachs-Institut, Julius-Maximilians-Universität Würzburg, Julius-von-Sachs Platz 2, 97082, Würzburg, Germany.
authors.forename: Nora
authors.initials: N
authors.lastname: Wehner
authors.affiliation: Pharmazeutische Biologie, Julius-von-Sachs-Institut, Julius-Maximilians-Universität Würzburg, Julius-von-Sachs Platz 2, 97082, Würzburg, Germany.
authors.forename: Jörn
authors.initials: J
authors.lastname: Herfert
authors.affiliation: Pharmazeutische Biologie, Julius-von-Sachs-Institut, Julius-Maximilians-Universität Würzburg, Julius-von-Sachs Platz 2, 97082, Würzburg, Germany. wolfgang.droege-laser@uni-wuerzburg.de.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Dröge-Laser
authors.affiliation: Pharmazeutische Biologie, Julius-von-Sachs-Institut, Julius-Maximilians-Universität Würzburg, Julius-von-Sachs Platz 2, 97082, Würzburg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Weiste
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D002458:Cell Fractionation; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant / Q000187:drug effects*; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D057166:High-Throughput Screening Assays; D007210:Indoleacetic Acids / Q000494:pharmacology*; D010937:Plant Growth Regulators / Q000494:pharmacology*; D011523:Protoplasts / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection / Q000379:methods
chemical_list: D007210:Indoleacetic Acids; D010937:Plant Growth Regulators; D014157:Transcription Factors
keywords: Auxin; Gene regulation; High throughput; Protoplast; Screening; Transcription factor