title: Capture Hybridization Analysis of DNA Targets.
abstract: There are numerous recent cases where chromatin modifying complexes associate with long noncoding RNA (lncRNA), stoking interest in lncRNA genomic localization and associated proteins. Capture Hybridization Analysis of RNA Targets (CHART) uses complementary oligonucleotides to purify an RNA with its associated genomic DNA or proteins from formaldehyde cross-linked chromatin. Deep sequencing of the purified DNA fragments gives a comprehensive profile of the potential lncRNA biological targets in vivo. The combined identification of the genomic localization of RNA and its protein partners can directly inform hypotheses about RNA function, including recruitment of chromatin modifying complexes. Here, we provide a detailed protocol on how to design antisense capture oligos and perform CHART in tissue culture cells.
authors.affiliation: Department of Molecular Biophysics & Biochemistry, Yale University, 27392, New Haven, CT, 06511, USA.|Chemical Biology Institute, Yale University, West Haven, CT, 06516, USA.
authors.forename: Alec N
authors.initials: AN
authors.lastname: Sexton
authors.affiliation: Department of Molecular Biophysics & Biochemistry, Yale University, 27392, New Haven, CT, 06511, USA.|Chemical Biology Institute, Yale University, West Haven, CT, 06516, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Machyna
authors.affiliation: Department of Molecular Biophysics & Biochemistry, Yale University, 27392, New Haven, CT, 06511, USA. matthew.simon@yale.edu.|Chemical Biology Institute, Yale University, West Haven, CT, 06516, USA. matthew.simon@yale.edu.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Simon
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000302:isolation & purification*; D042002:Chromatin Assembly and Disassembly / Q000235:genetics; D053938:DNA Fragmentation; D016678:Genome; D023281:Genomics / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009841:Oligonucleotides / Q000235:genetics; D062085:RNA, Long Noncoding / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D002843:Chromatin; D009841:Oligonucleotides; D062085:RNA, Long Noncoding
keywords: CHART; Chromatin; Genomic DNA; High-throughput sequencing; RNA; lincRNA; lncRNA