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Nat Commun ; 11(1): 1018, 2020 02 24.
Article de Anglais | MEDLINE | ID: mdl-32094342

RÉSUMÉ

Mammalian genomes encode tens of thousands of noncoding RNAs. Most noncoding transcripts exhibit nuclear localization and several have been shown to play a role in the regulation of gene expression and chromatin remodeling. To investigate the function of such RNAs, methods to massively map the genomic interacting sites of multiple transcripts have been developed; however, these methods have some limitations. Here, we introduce RNA And DNA Interacting Complexes Ligated and sequenced (RADICL-seq), a technology that maps genome-wide RNA-chromatin interactions in intact nuclei. RADICL-seq is a proximity ligation-based methodology that reduces the bias for nascent transcription, while increasing genomic coverage and unique mapping rate efficiency compared with existing methods. RADICL-seq identifies distinct patterns of genome occupancy for different classes of transcripts as well as cell type-specific RNA-chromatin interactions, and highlights the role of transcription in the establishment of chromatin structure.


Sujet(s)
Chromatine/métabolisme , Cartographie chromosomique/méthodes , Séquençage nucléotidique à haut débit/méthodes , ARN non traduit/génétique , Analyse de séquence d'ARN/méthodes , Animaux , Lignée cellulaire , Noyau de la cellule/génétique , Noyau de la cellule/métabolisme , Chromatine/génétique , Assemblage et désassemblage de la chromatine/génétique , Banque de gènes , Souris , Cellules souches embryonnaires de souris , ARN non traduit/métabolisme , Transcription génétique
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