title: Chromosome conformation capture on chip in single Drosophila melanogaster tissues.
abstract: Chromosomes are protein-DNA complexes that encode life. In a cell nucleus, chromosomes are folded in a highly specific manner, which connects strongly to some of their paramount functions, such as DNA replication and gene transcription. Chromosome conformation capture methodologies allow researchers to detect chromosome folding, by quantitatively measuring which genomic sequences are in close proximity in nuclear space. Here, we describe a modified chromosome conformation capture on chip (4C) protocol, which is specifically designed for detection of chromosome folding in a single Drosophila melanogaster tissue. Our protocol enables 4C analyses on a limited number of cells, which is crucial for fly tissues, because these contain relatively low numbers of cells. We used this protocol to demonstrate that target genes of Polycomb group proteins interact with each other in nuclear space of third instar larval brain cells. Major benefits of using D. melanogaster in 4C studies are: (1) powerful and tractable genetic approaches can be incorporated; (2) short generation time allows use of complex genotypes; and (3) compact and well annotated genome. We anticipate that our sensitized 4C method will be generally applicable to detect chromosome folding in other fly tissues.
authors.affiliation: Division of Molecular Genetics and the Centre for Biomedical Genetics, Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. b.tolhuis@nki.nl
authors.forename: Bas
authors.initials: B
authors.lastname: Tolhuis
authors.forename: Marleen
authors.initials: M
authors.lastname: Blom
authors.forename: Maarten
authors.initials: M
authors.lastname: van Lohuizen
mesh_terms: D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D059006:Chromosomes, Insect / Q000235:genetics*; D003627:Data Interpretation, Statistical; D004210:Dissection; D004331:Drosophila melanogaster / Q000235:genetics*; D004843:Epistasis, Genetic; D005663:Furans; D017344:Genes, Insect; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D020411:Oligonucleotide Array Sequence Analysis; D013876:Thiophenes; D016707:Tissue Fixation
chemical_list: C516387:3,4-bis-(2,4,5-trimethyl-thiophen-3-yl)furan-2,5-dione; D005663:Furans; D013876:Thiophenes