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Quantitative analysis of protein-RNA interactions in fission yeast
Characterizing the interactions between RNAs and proteins in vivo is key to better understand how organisms regulate gene expression. Here, we describe a robust and quantitative protocol to measure specific RNA-protein interactions in a native context using RNA immunoprecipitation (RIP). We provide...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9108979/ https://www.ncbi.nlm.nih.gov/pubmed/35586315 http://dx.doi.org/10.1016/j.xpro.2022.101373 |
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author | Elías-Villalobos, Alberto Duncan, Caia Mata, Juan Helmlinger, Dominique |
author_facet | Elías-Villalobos, Alberto Duncan, Caia Mata, Juan Helmlinger, Dominique |
author_sort | Elías-Villalobos, Alberto |
collection | PubMed |
description | Characterizing the interactions between RNAs and proteins in vivo is key to better understand how organisms regulate gene expression. Here, we describe a robust and quantitative protocol to measure specific RNA-protein interactions in a native context using RNA immunoprecipitation (RIP). We provide a comprehensive experimental framework to detect cotranslational interactions and detail the quantitative analysis of purified RNAs by PCR and high-throughput sequencing. Although we developed the protocol in fission yeast, it can be readily implemented in other yeast species. For complete details on the use and execution of this protocol, please refer to Toullec et al. (2021). |
format | Online Article Text |
id | pubmed-9108979 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-91089792022-05-17 Quantitative analysis of protein-RNA interactions in fission yeast Elías-Villalobos, Alberto Duncan, Caia Mata, Juan Helmlinger, Dominique STAR Protoc Protocol Characterizing the interactions between RNAs and proteins in vivo is key to better understand how organisms regulate gene expression. Here, we describe a robust and quantitative protocol to measure specific RNA-protein interactions in a native context using RNA immunoprecipitation (RIP). We provide a comprehensive experimental framework to detect cotranslational interactions and detail the quantitative analysis of purified RNAs by PCR and high-throughput sequencing. Although we developed the protocol in fission yeast, it can be readily implemented in other yeast species. For complete details on the use and execution of this protocol, please refer to Toullec et al. (2021). Elsevier 2022-05-09 /pmc/articles/PMC9108979/ /pubmed/35586315 http://dx.doi.org/10.1016/j.xpro.2022.101373 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Elías-Villalobos, Alberto Duncan, Caia Mata, Juan Helmlinger, Dominique Quantitative analysis of protein-RNA interactions in fission yeast |
title | Quantitative analysis of protein-RNA interactions in fission yeast |
title_full | Quantitative analysis of protein-RNA interactions in fission yeast |
title_fullStr | Quantitative analysis of protein-RNA interactions in fission yeast |
title_full_unstemmed | Quantitative analysis of protein-RNA interactions in fission yeast |
title_short | Quantitative analysis of protein-RNA interactions in fission yeast |
title_sort | quantitative analysis of protein-rna interactions in fission yeast |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9108979/ https://www.ncbi.nlm.nih.gov/pubmed/35586315 http://dx.doi.org/10.1016/j.xpro.2022.101373 |
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