Long intronic GAA*TTC repeats induce epigenetic changes and reporter gene silencing in a molecular model of Friedreich ataxia

Nucleic Acids Res. 2008 Nov;36(19):6056-65. doi: 10.1093/nar/gkn604. Epub 2008 Sep 27.

Abstract

Friedreich ataxia (FRDA) is caused by hyperexpansion of GAA*TTC repeats located in the first intron of the FXN gene, which inhibits transcription leading to the deficiency of frataxin. The FXN gene is an excellent target for therapeutic intervention since (i) 98% of patients carry the same type of mutation, (ii) the mutation is intronic, thus leaving the FXN coding sequence unaffected and (iii) heterozygous GAA*TTC expansion carriers with approximately 50% decrease of the frataxin are asymptomatic. The discovery of therapeutic strategies for FRDA is hampered by a lack of appropriate molecular models of the disease. Herein, we present the development of a new cell line as a molecular model of FRDA by inserting 560 GAA*TTC repeats into an intron of a GFP reporter minigene. The GFP_(GAA*TTC)(560) minigene recapitulates the molecular hallmarks of the mutated FXN gene, i.e. inhibition of transcription of the reporter gene, decreased levels of the reporter protein and hypoacetylation and hypermethylation of histones in the vicinity of the repeats. Additionally, selected histone deacetylase inhibitors, known to stimulate the FXN gene expression, increase the expression of the GFP_(GAA*TTC)(560) reporter. This FRDA model can be adapted to high-throughput analyses in a search for new therapeutics for the disease.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • DNA Repeat Expansion*
  • Frataxin
  • Friedreich Ataxia / genetics*
  • Gene Silencing*
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Heterochromatin / metabolism
  • Histones / metabolism
  • Humans
  • Introns
  • Iron-Binding Proteins / genetics*
  • Models, Genetic
  • Models, Molecular
  • Transcription, Genetic

Substances

  • Heterochromatin
  • Histones
  • Iron-Binding Proteins
  • Green Fluorescent Proteins