Modular and Molecular Optimization of a LOV (Light-Oxygen-Voltage)-Based Optogenetic Switch in Yeast

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dc.contributor.authorRomero, Andres
dc.contributor.authorRojas, Vicente
dc.contributor.authorDelgado, Veronica
dc.contributor.authorSalinas, Francisco
dc.contributor.authorLarrondo, Luis F.
dc.date.accessioned2025-01-20T22:09:47Z
dc.date.available2025-01-20T22:09:47Z
dc.date.issued2021
dc.description.abstractOptogenetic switches allow light-controlled gene expression with reversible and spatiotemporal resolution. In Saccharomyces cerevisiae, optogenetic tools hold great potential for a variety of metabolic engineering and biotechnology applications. In this work, we report on the modular optimization of the fungal light-oxygen-voltage (FUN-LOV) system, an optogenetic switch based on photoreceptors from the fungus Neurospora crassa. We also describe new switch variants obtained by replacing the Gal4 DNA-binding domain (DBD) of FUN-LOV with nine different DBDs from yeast transcription factors of the zinc cluster family. Among the tested modules, the variant carrying the Hap1p DBD, which we call "HAP-LOV", displayed higher levels of luciferase expression upon induction compared to FUN-LOV. Further, the combination of the Hap1p DBD with either p65 or VP16 activation domains also resulted in higher levels of reporter expression compared to the original switch. Finally, we assessed the effects of the plasmid copy number and promoter strength controlling the expression of the FUN-LOV and HAP-LOV components, and observed that when low-copy plasmids and strong promoters were used, a stronger response was achieved in both systems. Altogether, we describe a new set of blue-light optogenetic switches carrying different protein modules, which expands the available suite of optogenetic tools in yeast and can additionally be applied to other systems.
dc.fuente.origenWOS
dc.identifier.doi10.3390/ijms22168538
dc.identifier.eissn1422-0067
dc.identifier.urihttps://doi.org/10.3390/ijms22168538
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/94347
dc.identifier.wosidWOS:000689140600001
dc.issue.numero16
dc.language.isoen
dc.revistaInternational journal of molecular sciences
dc.rightsacceso restringido
dc.subjectyeast
dc.subjectoptogenetics
dc.subjectblue-light switch
dc.subjectgene expression
dc.titleModular and Molecular Optimization of a LOV (Light-Oxygen-Voltage)-Based Optogenetic Switch in Yeast
dc.typeartículo
dc.volumen22
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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