Automated Calibration of Optoplate LEDs to Reduce Light Dose Variation in Optogenetic Experiments

Grødem, Edvard Os; Sweeney, Kieran; McClean, Megan N.

doi:10.2144/btn-2020-0077

Cited by 11 publications

(12 citation statements)

References 5 publications

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“…The LPA is a published optogenetic tool that provides programmed illumination to each well of a 24-well plate. We assembled our LPA as described in reference 87 and calibrated as previously described ( 88 , 89 ).…”

Section: Methodsmentioning

confidence: 99%

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Optogenetic Tools for Control of Public Goods in Saccharomyces cerevisiae

Morales

Patel

Stewart

et al. 2021

mSphere

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Section: Methodsmentioning

confidence: 99%

“…S120VC; Thorlabs) and power meter (catalog no. PM100D; Thorlabs) as previously described ( 43 , 88 , 89 ).…”

Section: Methodsmentioning

confidence: 99%

Optogenetic Tools for Control of Public Goods in Saccharomyces cerevisiae

Morales

Patel

Stewart

et al. 2021

mSphere

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“…The Light Plate Apparatus (LPA) is a published optogenetic tool which provides programmed illumination to each well of a 24-well plate. We assembled our LPA as described in 87 and calibrated as previously described 88,89 .…”

Section: Methodsmentioning

confidence: 99%

“…For all illumination methods, response was assessed by flow (traditional or imaging) cytometry as described below or measurement of optical density. The light output of all light sources was measured and validated with a standard photodiode power sensor (Thorlabs, #S120VC) and power meter (Thorlabs, #PM100D) as previously described 43,88,89 .…”

Section: Methodsmentioning

confidence: 99%

Optogenetic tools for public goods control in Saccharomyces cerevisiae

Nm¹,

Cj³

et al. 2021

Preprint

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Microorganisms live in dense and diverse communities, with interactions between cells guiding community development and phenotype. The ability to perturb specific intercellular interactions in space and time provides a powerful route to determining the critical interactions and design rules for microbial communities. Approaches using optogenetic tools to modulate these interactions offer promise, as light can be exquisitely controlled in space and time. We report new plasmids for rapid integration of an optogenetic system into Saccharomyces cerevisiae to engineer light-control of expression of a gene of interest. In a proof-of-principle study, we demonstrate the ability to control a model cooperative interaction, namely the expression of the enzyme invertase (SUC2) which allows S. cerevisiae to hydrolyze sucrose and utilize it as a carbon source. We demonstrate that the strength of this cooperative interaction can be tuned in space and time by modulating light intensity and through spatial control of illumination. Spatial control of light allows cooperators and cheaters to be spatially segregated, and we show that the interplay between cooperative and inhibitory interactions in space can lead to pattern formation. Our strategy can be applied to achieve spatiotemporal control of expression of a gene of interest in Saccharomyces cerevisiae to perturb both intercellular and interspecies interactions.

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“…For instance, the light activation at variable amplitudes (LAVA) equipment has been successfully assembled and implemented to assess complex patterns of cell differentiation in human embryonic stem cells (Repina et al, 2020). Similarly, the optoPlate-96 allows simultaneous multichromatic stimulation under micro-cultivation conditions without cross-illumination between microwells (Bugaj & Lim, 2019;Grødem, Sweeney, & McClean, 2020;Thomas, Hörner, & Weber, 2020). Importantly, some of these devices have already been used to perform optogenetics in yeast, such as the Light Plate Apparatus (LPA).…”

Section: Optogenetic Switches For Yeast Biotechnologymentioning

confidence: 99%

The rise and shine of yeast optogenetics

Figueroa

Rojas

Romero

et al. 2020

Yeast

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Optogenetics refers to the control of biological processes with light. The activation of cellular phenomena by defined wavelengths has several advantages compared with traditional chemically inducible systems, such as spatiotemporal resolution, dose-response regulation, low cost, and moderate toxic effects. Optogenetics has been successfully implemented in yeast, a remarkable biological platform that is not only a model organism for cellular and molecular biology studies, but also a microorganism with diverse biotechnological applications. In this review, we summarize the main optogenetic systems implemented in the budding yeast Saccharomyces cerevisiae, which allow orthogonal control (by light) of gene expression, protein subcellular localization, reconstitution of protein activity, and protein sequestration by oligomerization. Furthermore, we review the application of optogenetic systems in the control of metabolic pathways, heterologous protein production and flocculation. We then revise an example of a previously described yeast optogenetic switch, named FUN-LOV, which allows precise and strong activation of the target gene. Finally, we describe optogenetic systems that have not yet been implemented in yeast, which could therefore be used to expand the panel of available tools in this biological chassis. In conclusion, a wide repertoire of optogenetic systems can be used to address fundamental biological questions and broaden the biotechnological toolkit in yeast. Take Away The budding yeast Saccharomyces cerevisiae has become a powerful biological platform for optogenetics, allowing the use of light to control diverse phenotypes. In this review, we summarized the main optogenetic tools implemented in yeast and their applications in metabolic engineering and biotechnology. Finally, we propose different sources of building blocks to expand the current repertoire of optogenetic systems in yeast.

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Automated Calibration of Optoplate LEDs to Reduce Light Dose Variation in Optogenetic Experiments

Cited by 11 publications

References 5 publications

Optogenetic Tools for Control of Public Goods in Saccharomyces cerevisiae

Optogenetic Tools for Control of Public Goods in Saccharomyces cerevisiae

Optogenetic tools for public goods control in Saccharomyces cerevisiae

The rise and shine of yeast optogenetics

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