Analysis of interactions between genomic loci through Chromosome Conformation Capture (3C)

• Published in: Current protocols in cell biology Vol. Chapter 22; p. Unit22.15
• Main Authors: El Kaderi, Belal, Medler, Scott, Ansari, Athar
• Format: Journal Article
• Language: English
• Published: United States 01.09.2012
• Subjects: Chromatin - chemistry; Chromatin - genetics; Chromatin - metabolism; Chromosomes, Fungal - chemistry; Chromosomes, Fungal - genetics; Chromosomes, Fungal - metabolism; Cross-Linking Reagents - chemistry; Densitometry - methods; Genetic Techniques; Nucleic Acid Conformation; Polymerase Chain Reaction - methods; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism
• ISSN: 1934-2616
• DOI: 10.1002/0471143030.cb2215s56

Genome architecture plays a significant role in the regulation of DNA-based cellular processes such as transcription and recombination. The successful accomplishment of these processes involves coordinated interaction of DNA elements located at a distance from each other. The 'Chromosome Conformation Capture' (3C) assay is a convenient tool for identification of physical association between spatially separated DNA elements in a cell under physiological conditions. The principle of 3C is to convert physical chromosomal interactions into specific DNA ligation products, which are then detected by PCR. The 3C protocol was originally used to identify long-range, stable chromosomal interactions in yeast. Here we describe a modified 3C procedure that can detect transient, short-range interactions of DNA elements separated by a distance of less than 700 bp. This method has been successfully used to detect dynamic interaction of transcription regulatory elements in yeast and can be used for detecting similar interactions of other genomic regions.