title: Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes.
abstract: A host of observations demonstrating the relationship between nuclear architecture and processes such as gene expression have led to a number of new technologies for interrogating chromosome positioning. Whereas some of these technologies reconstruct intermolecular interactions, others have enhanced our ability to visualize chromosomes in situ. Here, we describe an oligonucleotide- and PCR-based strategy for fluorescence in situ hybridization (FISH) and a bioinformatic platform that enables this technology to be extended to any organism whose genome has been sequenced. The oligonucleotide probes are renewable, highly efficient, and able to robustly label chromosomes in cell culture, fixed tissues, and metaphase spreads. Our method gives researchers precise control over the sequences they target and allows for single and multicolor imaging of regions ranging from tens of kilobases to megabases with the same basic protocol. We anticipate this technology will lead to an enhanced ability to visualize interphase and metaphase chromosomes.
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Beliveau
authors.forename: Eric F
authors.initials: EF
authors.lastname: Joyce
authors.forename: Nicholas
authors.initials: N
authors.lastname: Apostolopoulos
authors.forename: Feyza
authors.initials: F
authors.lastname: Yilmaz
authors.forename: Chamith Y
authors.initials: CY
authors.lastname: Fonseka
authors.forename: Ruth B
authors.initials: RB
authors.lastname: McCole
authors.forename: Yiming
authors.initials: Y
authors.lastname: Chang
authors.forename: Jin Billy
authors.initials: JB
authors.lastname: Li
authors.forename: Tharanga Niroshini
authors.initials: TN
authors.lastname: Senaratne
authors.forename: Benjamin R
authors.initials: BR
authors.lastname: Williams
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Rouillard
authors.forename: Chao-ting
authors.initials: CT
authors.lastname: Wu
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism; D020223:Chromosome Painting / Q000379:methods*; D002875:Chromosomes / Q000235:genetics; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics; D005260:Female; D015723:Gene Library; D016678:Genome / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase / Q000235:genetics; D008677:Metaphase / Q000235:genetics; D051379:Mice; D015345:Oligonucleotide Probes / Q000378:metabolism*; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D015345:Oligonucleotide Probes