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Quantitative analysis of protein-RNA interactions in fission yeast.

ABSTRACT: 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).

SUBMITTER: Elias-Villalobos A

PROVIDER: S-EPMC9108979 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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Quantitative analysis of protein-RNA interactions in fission yeast.

Elías-Villalobos Alberto A Duncan Caia C Mata Juan J Helmlinger Dominique D

STAR protocols 20220509 2

Characterizing the interactions between RNAs and proteins <i>in vivo</i> 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 ...[more]

PMID: 35586315

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