title: Optogenetics in Plants: Red/Far-Red Light Control of Gene Expression.
abstract: 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.
authors.affiliation: Institute of Synthetic Biology, University of Düsseldorf, Universitätsstrasse 1, 40225, Düsseldorf, Germany.|iGRAD Plant International Graduate Program for Plant Science, University of Düsseldorf, Düsseldorf, Germany.
authors.forename: Rocio
authors.initials: R
authors.lastname: Ochoa-Fernandez
authors.affiliation: Institute of Synthetic Biology, University of Düsseldorf, Universitätsstrasse 1, 40225, Düsseldorf, Germany.|Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Alberstrasse 19a, 79104, Freiburg, Germany.
authors.forename: Sophia L
authors.initials: SL
authors.lastname: Samodelov
authors.affiliation: Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104, Freiburg, Germany.
authors.forename: Simon M
authors.initials: SM
authors.lastname: Brandl
authors.affiliation: Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104, Freiburg, Germany.
authors.forename: Elke
authors.initials: E
authors.lastname: Wehinger
authors.affiliation: Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104, Freiburg, Germany.|Novartis Pharma AG, Biologics Process R&D, 4002, Basel, Switzerland.
authors.forename: Konrad
authors.initials: K
authors.lastname: Müller
authors.affiliation: Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Alberstrasse 19a, 79104, Freiburg, Germany.|Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104, Freiburg, Germany.|BIOSS - Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 18, 79104, Freiburg, Germany.
authors.forename: Wilfried
authors.initials: W
authors.lastname: Weber
authors.affiliation: Institute of Synthetic Biology, University of Düsseldorf, Universitätsstrasse 1, 40225, Düsseldorf, Germany. matias.zurbriggen@uni-duesseldorf.de.|Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104, Freiburg, Germany. matias.zurbriggen@uni-duesseldorf.de.
authors.forename: Matias D
authors.initials: MD
authors.lastname: Zurbriggen
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000528:radiation effects*; D008027:Light; D008163:Luminescent Measurements / Q000379:methods; D062308:Optogenetics / Q000379:methods*; D051939:Phytochrome B / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011523:Protoplasts / Q000378:metabolism / Q000528:radiation effects; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects
chemical_list: D029681:Arabidopsis Proteins; D051792:Basic Helix-Loop-Helix Proteins; D051939:Phytochrome B; D010940:Plant Proteins; C497730:PIF6 protein, Arabidopsis
keywords: Arabidopsis thaliana; Nicotiana tabacum; Plant leaf protoplasts; Plant optogenetics; Plant synthetic biology; Red light-inducible gene expression system