Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labeling
Summary: Translational regulation is a fundamental step in gene expression with critical roles in biological processes within a cell. Here, we describe a protocol to assess translation activity in mammalian cells by incorporation of O-propargyl-puromycin (OP-Puro). OP-Puro is a puromycin analog that...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722005342 |
| _version_ | 1817999671903125504 |
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| author | Jack Chun-Chieh Hsu Joanna B. Pawlak Maudry Laurent-Rolle Peter Cresswell |
| author_facet | Jack Chun-Chieh Hsu Joanna B. Pawlak Maudry Laurent-Rolle Peter Cresswell |
| author_sort | Jack Chun-Chieh Hsu |
| collection | DOAJ |
| description | Summary: Translational regulation is a fundamental step in gene expression with critical roles in biological processes within a cell. Here, we describe a protocol to assess translation activity in mammalian cells by incorporation of O-propargyl-puromycin (OP-Puro). OP-Puro is a puromycin analog that is incorporated into newly synthesized proteins and is detected by click chemistry reaction. We use OP-Puro labeling to assess translation activity between different cell types or cells under different growth conditions by confocal microscopy and flow cytometry.For complete details on the use and execution of this protocol, please refer to Hsu et al. (2021) and Hsu et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-14T03:12:05Z |
| format | Article |
| id | doaj.art-f19999c1b7c445e0aca9c0f1df622408 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-14T03:12:05Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f19999c1b7c445e0aca9c0f1df6224082022-12-22T02:15:34ZengElsevierSTAR Protocols2666-16672022-09-0133101654Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labelingJack Chun-Chieh Hsu0Joanna B. Pawlak1Maudry Laurent-Rolle2Peter Cresswell3Department of Immunobiology, Yale University School of Medicine, New Haven, CT 06520, USA; Corresponding authorDepartment of Immunobiology, Yale University School of Medicine, New Haven, CT 06520, USA; Section of Infectious Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USASection of Infectious Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USA; Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06520, USA; Center for Infection and Immunity, Yale University School of Medicine, New Haven, CT 06520, USADepartment of Immunobiology, Yale University School of Medicine, New Haven, CT 06520, USA; Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA; Corresponding authorSummary: Translational regulation is a fundamental step in gene expression with critical roles in biological processes within a cell. Here, we describe a protocol to assess translation activity in mammalian cells by incorporation of O-propargyl-puromycin (OP-Puro). OP-Puro is a puromycin analog that is incorporated into newly synthesized proteins and is detected by click chemistry reaction. We use OP-Puro labeling to assess translation activity between different cell types or cells under different growth conditions by confocal microscopy and flow cytometry.For complete details on the use and execution of this protocol, please refer to Hsu et al. (2021) and Hsu et al. (2022). : 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/S2666166722005342Cell biologyMolecular biologyGene expressionMolecular/Chemical probes |
| spellingShingle | Jack Chun-Chieh Hsu Joanna B. Pawlak Maudry Laurent-Rolle Peter Cresswell Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labeling STAR Protocols Cell biology Molecular biology Gene expression Molecular/Chemical probes |
| title | Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labeling |
| title_full | Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labeling |
| title_fullStr | Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labeling |
| title_full_unstemmed | Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labeling |
| title_short | Protocol for assessing translational regulation in mammalian cell lines by OP-Puro labeling |
| title_sort | protocol for assessing translational regulation in mammalian cell lines by op puro labeling |
| topic | Cell biology Molecular biology Gene expression Molecular/Chemical probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722005342 |
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