<i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact?
Integrating fluorescent genes including e<i>GFP</i> in the yeast genome is common practice for various applications, including cell visualization and population monitoring. The transformation of a commercial <i>S. cerevisiae</i> strain by integrating a cassette including a ge...
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| Format: | Article |
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MDPI AG
2022-04-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/10/4/781 |
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| author | Fanny Bordet Rémy Romanet Camille Eicher Cosette Grandvalet Géraldine Klein Régis Gougeon Anne Julien-Ortiz Chloé Roullier-Gall Hervé Alexandre |
| author_facet | Fanny Bordet Rémy Romanet Camille Eicher Cosette Grandvalet Géraldine Klein Régis Gougeon Anne Julien-Ortiz Chloé Roullier-Gall Hervé Alexandre |
| author_sort | Fanny Bordet |
| collection | DOAJ |
| description | Integrating fluorescent genes including e<i>GFP</i> in the yeast genome is common practice for various applications, including cell visualization and population monitoring. The transformation of a commercial <i>S. cerevisiae</i> strain by integrating a cassette including a gene encoding an EGFP protein in the <i>HO</i> gene was carried out using CRISPR-Cas9 technology. Although this type of integration is often used and described as neutral at the phenotypic level of the cell, we have highlighted that under alcoholic fermentation (in a Chardonnay must), it has an impact on the exometabolome. We observed 41 and 82 unique biomarkers for the S3 and S3GFP strains, respectively, as well as 28 biomarkers whose concentrations varied significantly between the wild-type and the modified strains. These biomarkers were mainly found to correspond to peptides. Despite similar phenotypic growth and fermentation parameters, high-resolution mass spectrometry allowed us to demonstrate, for the first time, that the peptidome is modified when integrating this cassette in the <i>HO</i> gene. |
| first_indexed | 2024-03-09T04:23:27Z |
| format | Article |
| id | doaj.art-f7241da7361f4adab7e3470da200b77b |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T04:23:27Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-f7241da7361f4adab7e3470da200b77b2023-12-03T13:45:06ZengMDPI AGMicroorganisms2076-26072022-04-0110478110.3390/microorganisms10040781<i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact?Fanny Bordet0Rémy Romanet1Camille Eicher2Cosette Grandvalet3Géraldine Klein4Régis Gougeon5Anne Julien-Ortiz6Chloé Roullier-Gall7Hervé Alexandre8Institut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceInstitut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceInstitut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceInstitut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceInstitut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceInstitut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceLallemand SAS, 19 Rue des Briquetiers, CEDEX, 31700 Blagnac, FranceInstitut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceInstitut Agro Dijon, PAM UMR A 02.102, Institut Universitaire de la Vigne et du Vin (IUVV), Université Bourgogne Franche-Comté, Rue Claude Ladrey, BP 27877, CEDEX, 21000 Dijon, FranceIntegrating fluorescent genes including e<i>GFP</i> in the yeast genome is common practice for various applications, including cell visualization and population monitoring. The transformation of a commercial <i>S. cerevisiae</i> strain by integrating a cassette including a gene encoding an EGFP protein in the <i>HO</i> gene was carried out using CRISPR-Cas9 technology. Although this type of integration is often used and described as neutral at the phenotypic level of the cell, we have highlighted that under alcoholic fermentation (in a Chardonnay must), it has an impact on the exometabolome. We observed 41 and 82 unique biomarkers for the S3 and S3GFP strains, respectively, as well as 28 biomarkers whose concentrations varied significantly between the wild-type and the modified strains. These biomarkers were mainly found to correspond to peptides. Despite similar phenotypic growth and fermentation parameters, high-resolution mass spectrometry allowed us to demonstrate, for the first time, that the peptidome is modified when integrating this cassette in the <i>HO</i> gene.https://www.mdpi.com/2076-2607/10/4/781<i>S. cerevisiae</i>eGFP<i>HO</i> geneCRISPR-Cas9 technologymetabolomics |
| spellingShingle | Fanny Bordet Rémy Romanet Camille Eicher Cosette Grandvalet Géraldine Klein Régis Gougeon Anne Julien-Ortiz Chloé Roullier-Gall Hervé Alexandre <i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact? Microorganisms <i>S. cerevisiae</i> eGFP <i>HO</i> gene CRISPR-Cas9 technology metabolomics |
| title | <i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact? |
| title_full | <i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact? |
| title_fullStr | <i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact? |
| title_full_unstemmed | <i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact? |
| title_short | <i>eGFP</i> Gene Integration in <i>HO</i>: A Metabolomic Impact? |
| title_sort | i egfp i gene integration in i ho i a metabolomic impact |
| topic | <i>S. cerevisiae</i> eGFP <i>HO</i> gene CRISPR-Cas9 technology metabolomics |
| url | https://www.mdpi.com/2076-2607/10/4/781 |
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