A modular yeast biosensor for low-cost point-of-care pathogen detection

Ostrov, Nili; Jiménez, Miguel; Billerbeck, Sonja; Brisbois, James; Matragrano, Joseph Antonio; Ager, Alastair; Cornish, Virginia W.

doi:10.1126/sciadv.1603221

Cited by 113 publications

(135 citation statements)

References 52 publications

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“…way components(Ostrov et al, 2017;Shaw et al, 2019) may not function as well in strains with altered sterol content.Importantly, recent studies have utilised the yeast pheromone response to separate yeast growth from product generation. The delay of production until cells have grown to a particular cell density (reviewed inVenayak, Anesiadis, Cluett, & Mahadevan, 2015) is advantageous if transgene expression and/or the heterologous product is toxic or if production severely starves the cells of metabolites required for growth Yu et al (2017).…”

mentioning

confidence: 99%

The wide‐ranging phenotypes of ergosterol biosynthesis mutants, and implications for microbial cell factories

Johnston

Moses

Rosser

2020

Yeast

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Yeast strains have been used extensively as robust microbial cell factories for the production of bulk and fine chemicals, including biofuels (bioethanol), complex pharmaceuticals (antimalarial drug artemisinin and opioid pain killers), flavours, and fragrances (vanillin, nootkatone, and resveratrol). In many cases, it is of benefit to suppress or modify ergosterol biosynthesis during strain engineering, for example, to increase thermotolerance or to increase metabolic flux through an alternate pathway.However, the impact of modifying ergosterol biosynthesis on engineered strains is discussed sparsely in literature, and little attention has been paid to the implications of these modifications on the general health and well-being of yeast. Importantly, yeast with modified sterol content exhibit a wide range of phenotypes, including altered organization and dynamics of plasma membrane, altered susceptibility to chemical treatment, increased tolerance to high temperatures, and reduced tolerance to other stresses such as high ethanol, salt, and solute concentrations. Here, we review the wide-ranging phenotypes of viable Saccharomyces cerevisiae strains with altered sterol content and discuss the implications of these for yeast as microbial cell factories.

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mentioning

confidence: 99%

The wide‐ranging phenotypes of ergosterol biosynthesis mutants, and implications for microbial cell factories

Johnston

Moses

Rosser

2020

Yeast

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“…As we have demonstrated here, our model strain also offers immediate applications for engineering yeast as biosensors, whether as single strains or as part of engineered consortia. Already efforts in synthetic biology have used engineered yeast biosensors as medical diagnostics 57 , for pathogen detection 58 , and as a tool for accelerating metabolic engineering 22,59 . In all these applications, it is desirable for the user to have control over the response to input and the magnitude of gene expression it triggers.…”

Section: Discussionmentioning

confidence: 99%

Engineering a model cell for rational tuning of GPCR signaling

Shaw

Yamauchi

Mead

et al. 2018

Preprint

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G protein-coupled receptor (GPCR) signaling is the primary method eukaryotes use to respond to specific cues in their environment. However, the relationship between stimulus and response for each GPCR is difficult to predict due to diversity in natural signal transduction architecture and expression.Using genome engineering in yeast, we here constructed an insulated, modular GPCR signal transduction system to study how the response to stimuli can be predictably tuned using synthetic tools. We delineated the contributions of a minimal set of key components via computational and experimental refactoring, identifying simple design principles for rationally tuning the dose-response.Using four different receptors, we demonstrate how this enables cells and consortia to be engineered to respond to desired concentrations of peptides, metabolites and hormones relevant to human health. This work enables rational tuning of cell sensing, while providing a framework to guide reprogramming of GPCR-based signaling in more complex systems.

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“…This contrasts with most previous solutions where the output signal is dependent on active cellular metabolism and/or gene expression for reporter protein production (Courbet et al 2015;Daringer et al 2014;Ostrov et al 2017). Lastly, the platform can potentially offer large cost savings to assay manufacturers as the cells autonomously express and surface display nanobodies which removes the need for the costly steps of antibody production, purification and attachment to latex particles surface.…”

Section: Bioassay Development Towards Use As a Real-world Diagnostic mentioning

confidence: 95%

A Low-Cost Biological Agglutination Assay for Medical Diagnostic Applications

Kylilis¹,

Riangrungroj²,

Lai³

et al. 2018

Preprint

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Affordable, easy-to-use diagnostic tests that can be readily deployed for point-of-care (POC) testing are key in addressing challenges in the diagnosis of medical conditions and for improving global health in general. Ideally, POC diagnostic tests should be highly selective for the biomarker, user-friendly, have a flexible design architecture and a low cost of production.Here we developed a novel agglutination assay based on whole E. coli cells surface-displaying nanobodies which bind selectively to a target protein analyte. As a proof-of-concept, we show the feasibility of this design as a new diagnostic platform by the detection of a model analyte at nanomolar concentrations. Moreover, we show that the design architecture is flexible by building assays optimized to detect a range of model analyte concentrations supported using straight-forward design rules and a mathematical model. Finally, we re-engineer E. coli cells for the detection of a medically relevant biomarker by the display of two different antibodies against the human fibrinogen and demonstrate a detection limit as low as 10 pM in diluted human plasma. Overall, we demonstrate that our agglutination technology fulfills the requirement of POC testing by combining low-cost nanobody production, customizable detection range and low detection limits. This technology has the potential to produce affordable diagnostics for both field-testing in the developing world, emergency or disaster relief sites as well as routine medical testing and personalized medicine. Keywords Whole-cell biosensor Synthetic biologyPoint-of-care (POC) testing Medical diagnosticsLatex agglutination test Immunoassay

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A modular yeast biosensor for low-cost point-of-care pathogen detection

Abstract: A yeast-based dipstick biosensor enables low-cost, on-site surveillance of pathogens.

Cited by 113 publications

References 52 publications

The wide‐ranging phenotypes of ergosterol biosynthesis mutants, and implications for microbial cell factories

The wide‐ranging phenotypes of ergosterol biosynthesis mutants, and implications for microbial cell factories

Engineering a model cell for rational tuning of GPCR signaling

A Low-Cost Biological Agglutination Assay for Medical Diagnostic Applications

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