Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiae
Summary: Here, we present a protocol for modulating the function of the Cth2 mRNA-binding protein (RBP) in Saccharomyces cerevisiae. We describe steps to amplify and integrate mutations in Cth2 that affect its stability and function. Next, we detail the functional assay to verify the activity of the...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723007748 |
| _version_ | 1797367784900919296 |
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| author | Praveen K. Patnaik Hanna Barlit Vyacheslav M. Labunskyy |
| author_facet | Praveen K. Patnaik Hanna Barlit Vyacheslav M. Labunskyy |
| author_sort | Praveen K. Patnaik |
| collection | DOAJ |
| description | Summary: Here, we present a protocol for modulating the function of the Cth2 mRNA-binding protein (RBP) in Saccharomyces cerevisiae. We describe steps to amplify and integrate mutations in Cth2 that affect its stability and function. Next, we detail the functional assay to verify the activity of the wild-type and mutant versions of Cth2 in yeast cells. This protocol can be adopted to modify the function of other RBPs with their respective functional mutations.For complete details on the use and execution of this protocol, please refer to Patnaik et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-08T17:23:06Z |
| format | Article |
| id | doaj.art-cced9913b5cb4bcf94de9bf16824bea1 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-08T17:23:06Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-cced9913b5cb4bcf94de9bf16824bea12024-01-03T04:12:58ZengElsevierSTAR Protocols2666-16672024-03-0151102807Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiaePraveen K. Patnaik0Hanna Barlit1Vyacheslav M. Labunskyy2Department of Dermatology, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118, USA; Corresponding authorDepartment of Dermatology, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118, USADepartment of Dermatology, Boston University Chobanian & Avedisian School of Medicine, Boston, MA 02118, USA; Corresponding authorSummary: Here, we present a protocol for modulating the function of the Cth2 mRNA-binding protein (RBP) in Saccharomyces cerevisiae. We describe steps to amplify and integrate mutations in Cth2 that affect its stability and function. Next, we detail the functional assay to verify the activity of the wild-type and mutant versions of Cth2 in yeast cells. This protocol can be adopted to modify the function of other RBPs with their respective functional mutations.For complete details on the use and execution of this protocol, please refer to Patnaik et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723007748Cell-based AssaysModel OrganismsMolecular BiologyGene Expression |
| spellingShingle | Praveen K. Patnaik Hanna Barlit Vyacheslav M. Labunskyy Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiae STAR Protocols Cell-based Assays Model Organisms Molecular Biology Gene Expression |
| title | Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiae |
| title_full | Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiae |
| title_fullStr | Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiae |
| title_full_unstemmed | Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiae |
| title_short | Manipulating mRNA-binding protein Cth2 function in budding yeast Saccharomyces cerevisiae |
| title_sort | manipulating mrna binding protein cth2 function in budding yeast saccharomyces cerevisiae |
| topic | Cell-based Assays Model Organisms Molecular Biology Gene Expression |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723007748 |
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