Engineered BRET-Based Biologic Light Sources Enable Spatiotemporal Control over Diverse Optogenetic Systems

• DOI: 10.1021/acssynbio.9b00277
• Publication Date: 2019-12-13T18:00:47Z
• Authors: Kshitij Parag-Sharma; Colin P. O’Banion; Erin C. Henry; Adele M. Musicant; John L. Cleveland; David S. Lawrence; Antonio L. Amelio
• Keywords: Bioluminescence Resonance Energy Transfer Techniques; 0301 basic medicine; Biological Products; Light; Green Fluorescent Proteins; Transfection; Optogenetics; Luminescent Proteins; 03 medical and health sciences; HEK293 Cells; Humans; Luciferases; HeLa Cells

Light-inducible optogenetic systems offer precise spatiotemporal control over a myriad of biologic processes. Unfortunately, current systems are inherently limited by their dependence on external light sources for their activation. Further, the utility of laser/LED-based illumination strategies are often constrained by the need for invasive surgical procedures to deliver such devices and local heat production, photobleaching and phototoxicity that compromises cell and tissue viability. To overcome these limitations, we developed a novel BRET-activated optogenetics (BEACON) system that employs biologic light to control optogenetic tools. BEACON is driven by self-illuminating bioluminescent-fluorescent proteins that generate "spectrally tuned" biologic light via bioluminescence resonance energy transfer (BRET). Notably, BEACON robustly activates a variety of commonly used optogenetic systems in a spatially restricted fashion, and at physiologically relevant time scales, to levels that are achieved by conventional laser/LED light sources.