Transcriptome-wide identification of RNA-binding protein and microRNA target sites by PAR-CLIP.
Cell
; 141(1): 129-41, 2010 Apr 02.
Article
en En
| MEDLINE
| ID: mdl-20371350
ABSTRACT
RNA transcripts are subject to posttranscriptional gene regulation involving hundreds of RNA-binding proteins (RBPs) and microRNA-containing ribonucleoprotein complexes (miRNPs) expressed in a cell-type dependent fashion. We developed a cell-based crosslinking approach to determine at high resolution and transcriptome-wide the binding sites of cellular RBPs and miRNPs. The crosslinked sites are revealed by thymidine to cytidine transitions in the cDNAs prepared from immunopurified RNPs of 4-thiouridine-treated cells. We determined the binding sites and regulatory consequences for several intensely studied RBPs and miRNPs, including PUM2, QKI, IGF2BP1-3, AGO/EIF2C1-4 and TNRC6A-C. Our study revealed that these factors bind thousands of sites containing defined sequence motifs and have distinct preferences for exonic versus intronic or coding versus untranslated transcript regions. The precise mapping of binding sites across the transcriptome will be critical to the interpretation of the rapidly emerging data on genetic variation between individuals and how these variations contribute to complex genetic diseases.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Técnicas Genéticas
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Proteínas de Unión al ARN
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ARN no Traducido
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MicroARNs
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Secuencias Reguladoras de Ácido Ribonucleico
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Límite:
Humans
Idioma:
En
Año:
2010
Tipo del documento:
Article