Using molecular tethering to analyze the role of nuclear compartmentalization in the regulation of mammalian gene activity

K. L. Reddy, H. Singh

Research output: Contribution to journalArticlepeer-review

Abstract

The mammalian nucleus has a complex structural organization that dynamically interacts with the genome. Chromatin is organized into discrete domains by association with distinct nuclear compartments enriched in structural and regulatory proteins. Growing evidence suggests that gene activity is modulated by interactions with these sub-nuclear compartments. Therefore, analyzing how nuclear architecture controls genome activity will be necessary to fully understand complex biological processes such as development and disease. In this article we describe a molecular methodology involving inducible tethering that can be used to position genes at the inner nuclear membrane (INM)-lamina compartment. The consequences of such directed re-positioning on gene activity or other DNA transactions can then be analyzed. This approach can be generalized and extended to position genes or chromosomal domains within other nuclear compartments thereby greatly facilitating the analysis of nuclear structure and its impact on genome activity.

Original languageEnglish (US)
Pages (from-to)242-251
Number of pages10
JournalMethods
Volume45
Issue number3
DOIs
StatePublished - Jul 2008
Externally publishedYes

Keywords

  • Confocal microscopy
  • Emerin
  • Immuno-FISH
  • Immunofluorescence
  • Inducible DNA tethering
  • Inner nuclear membrane
  • LacO
  • Lamin B
  • Nuclear compartmentalization
  • Nuclear lamina
  • Nuclear protein tether
  • Nuclear structure and gene activity
  • Re-positioning chromosomes
  • Targeting genes to nuclear compartments
  • lacI
  • lap2

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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