A detailed protocol for RNA cleavage assay in sympathetic neurons

This protocol illustrates the use of an in vitro assay to study the cleavage of the IMPA1 3′UTR by the endonuclease Ago2 in sympathetic neurons. The procedure includes the preparation of cytoplasmic protein extracts and also describes the synthesis and labeling of the RNA probe. The protocol can be...

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Detalles Bibliográficos
Autores principales: Andreassi, Catia, Riccio, Antonella
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672046/
https://www.ncbi.nlm.nih.gov/pubmed/34950884
http://dx.doi.org/10.1016/j.xpro.2021.101001
Descripción
Sumario:This protocol illustrates the use of an in vitro assay to study the cleavage of the IMPA1 3′UTR by the endonuclease Ago2 in sympathetic neurons. The procedure includes the preparation of cytoplasmic protein extracts and also describes the synthesis and labeling of the RNA probe. The protocol can be applied to other cell systems, RNA transcripts, and endonucleases to confirm the role of known cleavage site(s) and cleavage proteins, or to investigate new ones. For complete details on the use and execution of this protocol, please refer to Andreassi et al. (2021).

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