Construction of a Multiwell Light-Induction Platform for Traceless Control of Gene Expression in Mammalian Cells
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. Mehr anzeigen
Persistenter Link
https://doi.org/10.3929/ethz-b-000464930Publikationsstatus
publishedHerausgeber(in)
Buchtitel
Photoswitching Proteins. Methods and ProtocolsZeitschrift / Serie
Methods in Molecular BiologyBand
Seiten / Artikelnummer
Verlag
Humana PressThema
Optogenetics; Inducible gene expression; Cell engineering; Mammalian cells; Synthetic biologyOrganisationseinheit
03694 - Fussenegger, Martin / Fussenegger, Martin
Förderung
785800 - Electrogenetics - Shaping Electrogenetic Interfaces for Closed-Loop Voltage-Controlled Gene Expression (EC)