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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...

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Detalles Bibliográficos
Autores principales: Elías-Villalobos, Alberto, Duncan, Caia, Mata, Juan, Helmlinger, Dominique
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
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).
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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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