Mapping regulatory elements by DNaseI hypersensitivity chip (DNase-Chip).
Historically, the simplest method to robustly identify active gene regulatory elements has been enzymatic digestion of nuclear DNA by nucleases such as DNaseI. Regions of extreme chromatin accessibility to DNaseI, commonly known as DNaseI hypersensitive sites, have been repeatedly shown to be markers for all types of active cis-acting regulatory elements, including promoters, enhancers, silencers, insulators, and locus control regions. However, the original classical method, which for over 25 years relied on Southern blot, was limited to studying only small regions of the genome. Here we describe the detailed protocol for DNase-chip, a high-throughput method that allows for a targeted or genome-wide identification of cis-acting gene regulatory elements.
Shibata
Yoichiro
Y
Institute for Genome Sciences & Policy, Duke University, Durham, NC, USA.
Crawford
Gregory E
GE
D002874:Chromosome Mapping / Q000295:instrumentation / Q000379:methods*; D003850:Deoxyribonuclease I / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D012045:Regulatory Sequences, Nucleic Acid
D003850:Deoxyribonuclease I