title: Orthogonal optogenetic triple-gene control in Mammalian cells.
abstract: Optogenetic gene switches allow gene expression control at an unprecedented spatiotemporal resolution. Recently, light-responsive transgene expression systems that are activated by UV-B, blue, or red light have been developed. These systems perform well on their own, but their integration into genetic networks has been hampered by the overlapping absorbance spectra of the photoreceptors. We identified a lack of orthogonality between UV-B and blue light-controlled gene expression as the bottleneck and employed a model-based approach that identified the need for a blue light-responsive gene switch that is insensitive to low-intensity light. Based on this prediction, we developed a blue light-responsive and rapidly reversible expression system. Finally, we employed this expression system to demonstrate orthogonality between UV-B, blue, and red/far-red light-responsive gene switches in a single mammalian cell culture. We expect this approach to enable the spatiotemporal control of gene networks and to expand the applications of optogenetics in synthetic biology.
authors.affiliation: Faculty of Biology, University of Freiburg , Schänzlestrasse 1, 79104 Freiburg, Germany.
authors.forename: Konrad
authors.initials: K
authors.lastname: Müller
authors.forename: Raphael
authors.initials: R
authors.lastname: Engesser
authors.forename: Jens
authors.initials: J
authors.lastname: Timmer
authors.forename: Matias D
authors.initials: MD
authors.lastname: Zurbriggen
authors.forename: Wilfried
authors.initials: W
authors.lastname: Weber
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005786:Gene Expression Regulation / Q000235:genetics*; D062308:Optogenetics / Q000379:methods*; D058615:Synthetic Biology / Q000379:methods*
keywords: LOV; gene expression; multichromatic; optogenetics; synthetic biology