Assessment of fluorescent protein candidates for multi-color flow cytometry analysis of Saccharomyces cerevisiae

Foncillas, Raquel Perruca; Davidsson, Johan Bo; Carlquist, Magnus; Gorwa-Grauslund, Marie-Françoise

doi:10.1016/j.btre.2022.e00735

Cited by 8 publications

(3 citation statements)

References 33 publications

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“…This opens the possibility of further development of on-line flow cytometry not only as a monitoring tool but also as a controlling tool, in a similar manner as for CO 2 control-loops [ 25 ]. Together with the recent design of compatible fluorescent proteins in S. cerevisiae [ 24 , 28 ], flow cytometry analysis also paves the way for multiple and concomitant phenotype analysis at the single- cell level.…”

Section: Discussionmentioning

confidence: 99%

Assessment of the TRX2p-yEGFP Biosensor to Monitor the Redox Response of an Industrial Xylose-Fermenting Saccharomyces cerevisiae Strain during Propagation and Fermentation

Foncillas

Sanchis-Sebastiá

Wallberg

et al. 2023

JoF

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The commercial production of bioethanol from lignocellulosic biomass such as wheat straw requires utilizing a microorganism that can withstand all the stressors encountered in the process while fermenting all the sugars in the biomass. Therefore, it is essential to develop tools for monitoring and controlling the cellular fitness during both cell propagation and sugar fermentation to ethanol. In the present study, on-line flow cytometry was adopted to assess the response of the biosensor TRX2p-yEGFP for redox imbalance in an industrial xylose-fermenting strain of Saccharomyces cerevisiae during cell propagation and the following fermentation of wheat-straw hydrolysate. Rapid and transient induction of the sensor was recorded upon exposure to furfural and wheat straw hydrolysate containing up to 3.8 g/L furfural. During the fermentation step, the induction rate of the sensor was also found to correlate to the initial ethanol production rate, highlighting the relevance of redox monitoring and the potential of the presented tool to assess the ethanol production rate in hydrolysates. Three different propagation strategies were also compared, and it was confirmed that pre-exposure to hydrolysate during propagation remains the most efficient method for high ethanol productivity in the following wheat-straw hydrolysate fermentations.

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Section: Discussionmentioning

confidence: 99%

Assessment of the TRX2p-yEGFP Biosensor to Monitor the Redox Response of an Industrial Xylose-Fermenting Saccharomyces cerevisiae Strain during Propagation and Fermentation

Foncillas

Sanchis-Sebastiá

Wallberg

et al. 2023

JoF

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“…The steady-state expression level is determined by both the synthesis rate and the degradationand-dilution rate, while the induction kinetics are solely determined by the degradation-anddilution rate (Rosenfeld et al, 2002). Fluorescent proteins are highly stable and have in vivo half-lives longer than 20 hours (Corish and Tyler-Smith, 1999;He et al, 2019;Perruca-Foncillas et al, 2022). In our experimental setup, given that the doubling time of proliferating S. pombe cells is around 3 hours (Petersen and Russell, 2016), the degradation-and-dilution rate of mECitrine should mainly be determined by its dilution through cell division.…”

Section: Induction Kinetics In Dividing Cellsmentioning

confidence: 99%

An improved tetracycline-inducible expression system for fission yeast

Lyu,

Yang,

Pan

et al. 2024

Preprint

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The ability to manipulate gene expression is valuable for elucidating gene function. In the fission yeastSchizosaccharomyces pombe, the most widely used regulatable expression system is thenmt1promoter and its two attenuated variants. However, these promoters have limitations, including a long lag, incompatibility with rich media, and unsuitability for non- dividing cells. Here, we present a tetracycline-inducible system free of these shortcomings. Our system features theenotetSpromoter, which achieves a similar induced level and a higher induction ratio compared to thenmt1promoter, without exhibiting a lag. Additionally, our system includes four weakenedenotetSvariants, offering an expression range similar to thenmt1series promoters but with more intermediate levels. To enhance usability, each promoter is combined with a Tet-repressor-expressing cassette in an integration plasmid. Importantly, our system can be used in non-dividing cells, enabling the development of a synchronous meiosis induction method with high spore viability. Moreover, our system allows for the shutdown of gene expression and the generation of conditional loss-of-function mutants. This system provides a versatile and powerful tool for manipulating gene expression in fission yeast.

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“…At present, chemiluminescence has replaced enzyme-linked immunoassay as the mainstream immunodiagnostic method; the market size has reached more than 70% of the total immunodiagnostic market and is basically monopolized by the leading companies. In the future, with the breakthrough of key core technologies and the introduction of advanced technologies, coupled with the support of policies to encourage innovation, important breakthroughs will be made in the field of chemiluminescence, which is expected to gradually realize leapfrog development in the field of high-end immunodiagnostics [84][85][86][87].…”

Section: Chemiluminescencementioning

confidence: 99%

Methods and Advances in the Design, Testing and Development of In Vitro Diagnostic Instruments

Wang

et al. 2023

Processes

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With the continuous improvement of medical testing and instrumentation engineering technologies, the design, testing and development methods of in vitro diagnostic instruments are developing rapidly. In vitro diagnostic instruments are also gradually developing into a class of typical high-end medical equipment. The design of in vitro diagnostic instruments involves a variety of medical diagnostic methods and biochemical, physical and other related technologies, and its development process involves complex system engineering. This paper systematically organizes and summarizes the design, testing and development methods of in vitro diagnostic instruments and their development in recent years, focusing on summarizing the related technologies and core aspects of the R&D process, and analyzes the development trend of the in vitro diagnostic instrument market.

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Assessment of fluorescent protein candidates for multi-color flow cytometry analysis of Saccharomyces cerevisiae

Cited by 8 publications

References 33 publications

Assessment of the TRX2p-yEGFP Biosensor to Monitor the Redox Response of an Industrial Xylose-Fermenting Saccharomyces cerevisiae Strain during Propagation and Fermentation

Assessment of the TRX2p-yEGFP Biosensor to Monitor the Redox Response of an Industrial Xylose-Fermenting Saccharomyces cerevisiae Strain during Propagation and Fermentation

An improved tetracycline-inducible expression system for fission yeast

Methods and Advances in the Design, Testing and Development of In Vitro Diagnostic Instruments

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