Optogenetics in Plants: Red/Far-Red Light Control of Gene Expression
Optogenetic tools to control gene expression have many advantages over the classical chemically inducible systems, overcoming intrinsic limitations of chemical inducers such as solubility, diffusion, and cell toxicity. They offer an unmatched spatiotemporal resolution and permit quantitative and noninvasive control of the gene expression. Here we describe a protocol of a synthetic light-inducible system for the targeted control of gene expression in plants based on the plant photoreceptor phytochrome B and one of its interacting factors (PIF6). The synthetic toggle switch system is in the ON state when plant protoplasts are illuminated with red light (660 nm) and can be returned to the OFF state by subsequent illumination with far-red light (760 nm). In this protocol, the implementation of a red light-inducible expression system in plants using Light-Emitting Diode (LED) illumination boxes is described, including the isolation and transient transformation of plant protoplasts from Arabidopsis thaliana and Nicotiana tabacum.
Key wordsPlant synthetic biology Plant optogenetics Red light-inducible gene expression system Plant leaf protoplasts Arabidopsis thaliana Nicotiana tabacum
This work was supported in part by the Excellence Initiative of the German Federal and State Governments (EXC294-BIOSS, GSC-4 Spemann Graduate School (SGBM)) and the Alexander von Humbolt Foundation (research Grant no. 1141629). We thank Susanne Knall and Frauke Bartels-Burgahn for experimental assistance. We thank J. Schmidt, D. Schächtele and J. Meßmer (University of Freiburg) for designing and constructing the illumination boxes.
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