Rounding up active cis-elements in the triple C corral: combining conservation, cleavage and conformation capture for the analysis of regulatory gene domains.
Identification and functional analysis of potential cis-regulatory elements is a laborious process that often depends on removing putative elements from their natural context to study their activity. While such methods provide valuable information about the isolated element, they disregard the potential role of an element's interaction(s) with other regulatory sequences and the three-dimensional structure of an active gene locus. Here, two novel methods are discussed--chromosome conformation capture (3C) and RNA-TRAP--that can be used to detect interactions between distal regulatory sites and which thus indicate the chromosomal conformation that is adopted by a gene locus in various states of transcriptional activity. Combined with comparative genomics and traditional DNase I hypersensitive site mapping, these methods form a powerful approach for the study of the mechanisms of long-range transcriptional regulation.
McBride
David J
DJ
Cell and Molecular Genetics Section, MRC Human Genetics Unit, Edinburgh, EH4 2XU, UK.
Kleinjan
Dirk A
DA
D000818:Animals; D002843:Chromatin / Q000378:metabolism; D002875:Chromosomes / Q000648:ultrastructure; D003850:Deoxyribonuclease I / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D023281:Genomics / Q000379:methods; D006801:Humans; D051379:Mice; D008957:Models, Genetic; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D040901:Proteomics; D012313:RNA / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid
D002843:Chromatin; D012313:RNA; D003850:Deoxyribonuclease I