Abstract
Understanding the context-specific role of gene function is a key objective of modern biology. To this end, we generated a resource for inducible cell type-specific transactivation in Arabidopsis (Arabidopsis thaliana) based on the well-established combination of the chimeric GR-LhG4 transcription factor and the synthetic pOp promoter. Harnessing the flexibility of the GreenGate cloning system, we produced a comprehensive set of transgenic lines termed GR-LhG4 driver lines targeting most tissues in the Arabidopsis shoot and root with a strong focus on the indeterminate meristems. When we combined these transgenic lines with effectors under the control of the pOp promoter, we observed tight temporal and spatial control of gene expression. In particular, inducible expression in F1 plants obtained from crosses of driver and effector lines allows for rapid assessment of the cell type-specific impact of an effector with high temporal resolution. Thus, our comprehensive and flexible method is suitable for overcoming the limitations of ubiquitous genetic approaches, the outputs of which often are difficult to interpret due to the widespread existence of compensatory mechanisms and the integration of diverging effects in different cell types.
Highlights
Ann-Kathrin Schürholz,2 Vadir López-Salmerón,2 Zhenni Li, Joachim Forner,3 Christian Wenzl, Christophe Gaillochet, Sebastian Augustin,4 Amaya Vilches Barro, Michael Fuchs, Michael Gebert, Jan U
The glucocorticoid receptor (GR)-LhG4 transcription factor is expressed under the control of a tissue- or cell type-specific promoter
We anticipate that the most utility can be obtained from this system if lines harboring effector cassettes are crossed with driver lines and analyses are performed with F1 plants
Summary
Understanding the context-specific role of gene function is a key objective of modern biology To this end, we generated a resource for inducible cell type-specific transactivation in Arabidopsis (Arabidopsis thaliana) based on the well-established combination of the chimeric GR-LhG4 transcription factor and the synthetic pOp promoter. Harnessing the flexibility of the GreenGate cloning system, we produced a comprehensive set of transgenic lines termed GR-LhG4 driver lines targeting most tissues in the Arabidopsis shoot and root with a strong focus on the indeterminate meristems When we combined these transgenic lines with effectors under the control of the pOp promoter, we observed tight temporal and spatial control of gene expression. Transgenic and mutational approaches can interfere with plant vitality, precluding an in-depth analysis Many of these problems can be overcome by inducible, cell type-specific expression mediated by twocomponent transcription activation systems (Moore et al, 2006). For the same reason and because distinct tissue-specific promoters were not always available in the past, attention is usually given to one tissue or cell type of interest at a time, and unbiased approaches targeting a larger spectrum of individual tissues are rarely followed
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