Mapping cis- and trans- chromatin interaction networks using chromosome conformation capture (3C).

摘要: Expression of genes can be controlled by regulatory elements that are located at large genomic distances from their target (in cis), or even on different chromosomes trans). Regulatory act engaging in direct physical interactions with resulting the formation chromatin loops. Thus, and come close spatial proximity irrespective relative positions. Analysis between will reveal which regulate each gene, provide fundamental insights into organization general. Long-range cis- trans- studied high resolution using chromosome conformation capture (3C) technology. 3C employs formaldehyde crosslinking to trap loci throughout genome. Crosslinked cells solubilized is digested a restriction enzyme. Chromatin subsequently ligated under conditions favor intramolecular ligation. After reversal crosslinks, DNA purified interaction frequencies specific chromosomal determined quantifying amounts corresponding ligation products polymerase chain reaction (PCR). This chapter describes detailed protocols for analysis yeast Saccharomyces cerevisiae mammalian cells.