Abstract
Mammalian cells can be engineered to incorporate light-responsive elements that reliably sense stimulation by light and activate endogenous pathways, such as the cAMP or Ca2+ pathway, to control gene expression. Light-inducible gene expression systems offer high spatiotemporal resolution, and are also traceless, reversible, tunable, and inexpensive. Melanopsin, a well-known representative of the animal opsins, is a G-protein-coupled receptor that triggers a Gαq-dependent signaling cascade upon activation with blue light (≈470 nm). Here, we describe how to rewire melanopsin activation by blue light to transgene expression in mammalian cells, with detailed instructions for constructing a 96-LED array platform with multiple tunable parameters for illumination of the engineered cells in multiwell plates.
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Acknowledgments
This work was supported by the European Research Council (ERC) advanced grant (ElectroGene; grant no. 785800) and in part by the National Centre of Competence in Research (NCCR) for Molecular Systems Engineering.
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Mansouri, M., Lichtenstein, S., Strittmatter, T., Buchmann, P., Fussenegger, M. (2020). Construction of a Multiwell Light-Induction Platform for Traceless Control of Gene Expression in Mammalian Cells. In: Niopek, D. (eds) Photoswitching Proteins . Methods in Molecular Biology, vol 2173. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0755-8_13
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DOI: https://doi.org/10.1007/978-1-0716-0755-8_13
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