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An Optimized Immunoprecipitation Protocol for Assessing Protein-RNA Interactions In Vitro.

ABSTRACT: RNA-binding proteins are key regulators of cell identity and function, which underscores the need for unbiased and versatile protocols to identify and characterize novel protein-RNA interactions. Here, we describe a simple and cost-effective in vitro RNA immunoprecipitation (iv-RIP) method to assess the direct or indirect RNA-binding ability of any protein of interest. The versatility of this method relies on the adaptability of the immunoprecipitation conditions and the choice of the RNA, which exponentially broadens the range of potential applications. For complete details on the use and execution of this protocol, please refer to Guallar et al. (2020).

SUBMITTER: Fuentes-Iglesias A

PROVIDER: S-EPMC7521669 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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An Optimized Immunoprecipitation Protocol for Assessing Protein-RNA Interactions <i>In Vitro</i>.

Fuentes-Iglesias Alejandro A Garcia-Outeiral Vera V Pardavila Jose Angel JA Wang Jianlong J Fidalgo Miguel M Guallar Diana D

STAR protocols 20200826 2

RNA-binding proteins are key regulators of cell identity and function, which underscores the need for unbiased and versatile protocols to identify and characterize novel protein-RNA interactions. Here, we describe a simple and cost-effective <i>in vitro</i> RNA immunoprecipitation (iv-RIP) method to assess the direct or indirect RNA-binding ability of any protein of interest. The versatility of this method relies on the adaptability of the immunoprecipitation conditions and the choice of the RNA ...[more]

PMID: 32995755

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