RNA-Binding Protein Immunoprecipitation and High-Throughput Sequencing.
Methods Mol Biol
; 2200: 453-461, 2021.
Article
em En
| MEDLINE
| ID: mdl-33175393
ABSTRACT
The RNA-binding proteome plays a key role in controlling every step in the life of RNA molecules. Through interaction with dedicated sequence motifs, RNA-binding proteins coordinate processing of cohorts of genes. Understanding such posttranscriptional networks controlled by an RNA-binding protein requires a comprehensive identification of its in vivo targets. In Arabidopsis thaliana, RNA immunoprecipitation followed by reverse transcription-PCR has been widely used to test the association of candidate targets with RNA-binding proteins. The detection of unknown target transcripts requires methods operating at the level of the entire transcriptome. Here, we describe a protocol for RNA immunoprecipitation coupled to the generation of libraries from the co-purified RNAs for high-throughput sequencing. This allows determining RNAs associated with RNA-binding proteins in planta at a global scale.
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MEDLINE
Assunto principal:
Proteínas de Ligação a RNA
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Arabidopsis
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RNA de Plantas
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Reação em Cadeia da Polimerase Via Transcriptase Reversa
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Proteínas de Arabidopsis
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Imunoprecipitação
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Sequenciamento de Nucleotídeos em Larga Escala
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article