A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells.
Growth and differentiation of multicellular systems is orchestrated by spatially restricted gene expression programs in specialized subpopulations. The targeted manipulation of such processes by synthetic tools with high-spatiotemporal resolution could, therefore, enable a deepened understanding of developmental processes and open new opportunities in tissue engineering. Here, we describe the first red/far-red light-triggered gene switch for mammalian cells for achieving gene expression control in time and space. We show that the system can reversibly be toggled between stable on- and off-states using short light pulses at 660 or 740 nm. Red light-induced gene expression was shown to correlate with the applied photon number and was compatible with different mammalian cell lines, including human primary cells. The light-induced expression kinetics were quantitatively analyzed by a mathematical model. We apply the system for the spatially controlled engineering of angiogenesis in chicken embryos. The system's performance combined with cell- and tissue-compatible regulating red light will enable unprecedented spatiotemporally controlled molecular interventions in mammalian cells, tissues and organisms.
Faculty of Biology, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, Germany.
Konrad
K
Müller
Raphael
R
Engesser
Stéphanie
S
Metzger
Simon
S
Schulz
Michael M
MM
Kämpf
Moritz
M
Busacker
Thorsten
T
Steinberg
Pascal
P
Tomakidi
Martin
M
Ehrbar
Ferenc
F
Nagy
Jens
J
Timmer
Matias D
MD
Zubriggen
Wilfried
W
Weber
D000818:Animals; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002642:Chick Embryo; D006224:Cricetinae; D005786:Gene Expression Regulation / Q000528:radiation effects*; D006801:Humans; D008027:Light; D051379:Mice; D018919:Neovascularization, Physiologic / Q000235:genetics / Q000528:radiation effects; D051939:Phytochrome B / Q000235:genetics / Q000378:metabolism; D019076:Transgenes
D029681:Arabidopsis Proteins; D051792:Basic Helix-Loop-Helix Proteins; D051939:Phytochrome B; C497952:PHYB protein, Arabidopsis; C497730:PIF6 protein, Arabidopsis