Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library
Abstract Background Acetic acid tolerance is crucial for the development of robust cell factories for conversion of lignocellulosic hydrolysates that typically contain high levels of acetic acid. Screening mutants for growth in medium with acetic acid is an attractive way to identify sensitive varia...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-10-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-022-01938-7 |
| _version_ | 1811182247405944832 |
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| author | Maurizio Mormino Ibai Lenitz Verena Siewers Yvonne Nygård |
| author_facet | Maurizio Mormino Ibai Lenitz Verena Siewers Yvonne Nygård |
| author_sort | Maurizio Mormino |
| collection | DOAJ |
| description | Abstract Background Acetic acid tolerance is crucial for the development of robust cell factories for conversion of lignocellulosic hydrolysates that typically contain high levels of acetic acid. Screening mutants for growth in medium with acetic acid is an attractive way to identify sensitive variants and can provide novel insights into the complex mechanisms regulating the acetic acid stress response. Results An acetic acid biosensor based on the Saccharomyces cerevisiae transcription factor Haa1, was used to screen a CRISPRi yeast strain library where dCas9-Mxi was set to individually repress each essential or respiratory growth essential gene. Fluorescence-activated cell sorting led to the enrichment of a population of cells with higher acetic acid retention. These cells with higher biosensor signal were demonstrated to be more sensitive to acetic acid. Biosensor-based screening of the CRISPRi library strains enabled identification of strains with increased acetic acid sensitivity: strains with gRNAs targeting TIF34, MSN5, PAP1, COX10 or TRA1. Conclusions This study demonstrated that biosensors are valuable tools for screening and monitoring acetic acid tolerance in yeast. Fine-tuning the expression of essential genes can lead to altered acetic acid tolerance. |
| first_indexed | 2024-04-11T09:29:24Z |
| format | Article |
| id | doaj.art-d70653317fc94c54be9bd4f73b67a0c3 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-11T09:29:24Z |
| publishDate | 2022-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-d70653317fc94c54be9bd4f73b67a0c32022-12-22T04:31:56ZengBMCMicrobial Cell Factories1475-28592022-10-0121111410.1186/s12934-022-01938-7Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi libraryMaurizio Mormino0Ibai Lenitz1Verena Siewers2Yvonne Nygård3Department of Biology and Biological Engineering, Chalmers University of TechnologyDepartment of Biology and Biological Engineering, Chalmers University of TechnologyDepartment of Biology and Biological Engineering, Chalmers University of TechnologyDepartment of Biology and Biological Engineering, Chalmers University of TechnologyAbstract Background Acetic acid tolerance is crucial for the development of robust cell factories for conversion of lignocellulosic hydrolysates that typically contain high levels of acetic acid. Screening mutants for growth in medium with acetic acid is an attractive way to identify sensitive variants and can provide novel insights into the complex mechanisms regulating the acetic acid stress response. Results An acetic acid biosensor based on the Saccharomyces cerevisiae transcription factor Haa1, was used to screen a CRISPRi yeast strain library where dCas9-Mxi was set to individually repress each essential or respiratory growth essential gene. Fluorescence-activated cell sorting led to the enrichment of a population of cells with higher acetic acid retention. These cells with higher biosensor signal were demonstrated to be more sensitive to acetic acid. Biosensor-based screening of the CRISPRi library strains enabled identification of strains with increased acetic acid sensitivity: strains with gRNAs targeting TIF34, MSN5, PAP1, COX10 or TRA1. Conclusions This study demonstrated that biosensors are valuable tools for screening and monitoring acetic acid tolerance in yeast. Fine-tuning the expression of essential genes can lead to altered acetic acid tolerance.https://doi.org/10.1186/s12934-022-01938-7Acetic acidBiosensorLibraryScreeningToleranceCRISPRi |
| spellingShingle | Maurizio Mormino Ibai Lenitz Verena Siewers Yvonne Nygård Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library Microbial Cell Factories Acetic acid Biosensor Library Screening Tolerance CRISPRi |
| title | Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library |
| title_full | Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library |
| title_fullStr | Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library |
| title_full_unstemmed | Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library |
| title_short | Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library |
| title_sort | identification of acetic acid sensitive strains through biosensor based screening of a saccharomyces cerevisiae crispri library |
| topic | Acetic acid Biosensor Library Screening Tolerance CRISPRi |
| url | https://doi.org/10.1186/s12934-022-01938-7 |
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