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

ABSTRACT

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.


Subject(s)
Chromatin/metabolism , Chromosome Mapping/methods , High-Throughput Nucleotide Sequencing/methods , RNA, Untranslated/genetics , Sequence Analysis, RNA/methods , Animals , Cell Line , Cell Nucleus/genetics , Cell Nucleus/metabolism , Chromatin/genetics , Chromatin Assembly and Disassembly/genetics , Gene Library , Mice , Mouse Embryonic Stem Cells , RNA, Untranslated/metabolism , Transcription, Genetic
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