| Literature DB >> 17179217 |
Paul G Giresi1, Jonghwan Kim, Ryan M McDaniell, Vishwanath R Iyer, Jason D Lieb.
Abstract
DNA segments that actively regulate transcription in vivo are typically characterized by eviction of nucleosomes from chromatin and are experimentally identified by their hypersensitivity to nucleases. Here we demonstrate a simple procedure for the isolation of nucleosome-depleted DNA from human chromatin, termed FAIRE (Formaldehyde-Assisted Isolation of Regulatory Elements). To perform FAIRE, chromatin is crosslinked with formaldehyde in vivo, sheared by sonication, and phenol-chloroform extracted. The DNA recovered in the aqueous phase is fluorescently labeled and hybridized to a DNA microarray. FAIRE performed in human cells strongly enriches DNA coincident with the location of DNaseI hypersensitive sites, transcriptional start sites, and active promoters. Evidence for cell-type-specific patterns of FAIRE enrichment is also presented. FAIRE has utility as a positive selection for genomic regions associated with regulatory activity, including regions traditionally detected by nuclease hypersensitivity assays.Entities:
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Year: 2006 PMID: 17179217 PMCID: PMC1891346 DOI: 10.1101/gr.5533506
Source DB: PubMed Journal: Genome Res ISSN: 1088-9051 Impact factor: 9.043