A novel single cell method to identify the genetic composition at a single nuclear body

Sci Rep. 2016 Jul 8:6:29191. doi: 10.1038/srep29191.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence / genetics
  • Binding Sites / genetics
  • Cell Compartmentation / genetics*
  • Cell Nucleus / genetics*
  • Gene Expression Regulation
  • HeLa Cells
  • Humans
  • Nuclear Envelope / genetics*
  • Single-Cell Analysis / methods*

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