Open AccessChromatin‐Associated RNA Sequencing (ChAR‐seq)
Author(s) -
Jukam David,
Limouse Charles,
Smith Owen K.,
Risca Viviana I.,
Bell Jason C.,
Straight Aaron F.
Publication year - 2019
Publication title -
current protocols in molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.533
H-Index - 42
eISSN - 1934-3647
pISSN - 1934-3639
DOI - 10.1002/cpmb.87
Subject(s) - chromatin , rna , biology , non coding rna , computational biology , dna , genetics , rna splicing , genome , long non coding rna , gene
RNA is a fundamental component of chromatin. Noncoding RNAs (ncRNAs) can associate with chromatin to influence gene expression and chromatin state; many also act at long distances from their transcriptional origin. Yet we know almost nothing about the functions or sites of action for most ncRNAs. Current methods to identify sites of RNA interaction with the genome are limited to the study of a single RNA at a time. Here we describe a protocol for ChAR‐seq, a strategy to identify all chromatin‐associated RNAs and map their DNA contacts genome‐wide. In ChAR‐seq, proximity ligation of RNA and DNA to a linker molecule is used to construct a chimeric RNA‐DNA molecule that is converted to DNA for sequencing. In a single assay, ChAR‐seq can discover de novo chromatin interactions of distinct RNAs, including nascent transcripts, splicing RNAs, and long noncoding RNAs (lncRNAs). Resulting “maps” of genome‐bound RNAs should provide new insights into RNA biology. © 2019 by John Wiley & Sons, Inc.