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

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...

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Detalles Bibliográficos
Autores principales: Fuentes-Iglesias, Alejandro, Garcia-Outeiral, Vera, Pardavila, Jose Angel, Wang, Jianlong, Fidalgo, Miguel, Guallar, Diana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7521669/
https://www.ncbi.nlm.nih.gov/pubmed/32995755
http://dx.doi.org/10.1016/j.xpro.2020.100093
Descripción
Sumario: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).