title: A novel single cell method to identify the genetic composition at a single nuclear body.
abstract: Gene loci make specific associations with compartments of the nucleus (e.g. the nuclear envelope, nucleolus, and transcription factories) and this association may determine or reflect a mechanism of genetic control. With current methods, it is not possible to identify sets of genes that converge to form a "gene hub" as there is a reliance on loci-specific probes, or immunoprecipitation of a particular protein from bulk cells. We introduce a method that will allow for the identification of loci contained within the vicinity of a single nuclear body in a single cell. For the first time, we demonstrate that the DNA sequences originating from a single sub-nuclear structure in a single cell targeted by two-photon irradiation can be determined, and mapped to a particular locus. Its application to single PML nuclear bodies reveals ontologically related loci that frequently associate with each other and with PML bodies in a population of cells, and a possible nuclear body targeting role for specific transcription factor binding sites.
authors.affiliation: Genetics and Genome Biology Program, The Hospital for Sick Children, Toronto M5G 0A4, Canada.
authors.forename: David
authors.initials: D
authors.lastname: Anchel
authors.affiliation: Max Planck Institute of Immunobiology and Epigenetics, Department of Epigenetics, Freiburg 79108, Germany.
authors.forename: Reagan W
authors.initials: RW
authors.lastname: Ching
authors.affiliation: Genetics and Genome Biology Program, The Hospital for Sick Children, Toronto M5G 0A4, Canada.
authors.forename: Rachel
authors.initials: R
authors.lastname: Cotton
authors.affiliation: Genetics and Genome Biology Program, The Hospital for Sick Children, Toronto M5G 0A4, Canada.
authors.forename: Ren
authors.initials: R
authors.lastname: Li
authors.affiliation: Genetics and Genome Biology Program, The Hospital for Sick Children, Toronto M5G 0A4, Canada.
authors.forename: David P
authors.initials: DP
authors.lastname: Bazett-Jones
mesh_terms: D001483:Base Sequence / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D002451:Cell Compartmentation / Q000235:genetics*; D002467:Cell Nucleus / Q000235:genetics*; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D009685:Nuclear Envelope / Q000235:genetics*; D059010:Single-Cell Analysis / Q000379:methods*