Neurogenetics pp 101-108 | Cite as

Fluorescence PCR and GeneScan® Analysis for the Detection of CAG Repeat Expansions Associated with Huntington’s Disease

  • Cindy L. Vnencak-Jones
Part of the Methods in Molecular Biology™ book series (MIMB, volume 217)

Abstract

Huntington’s Disease (HD) is an autosomal dominant neurodegenerative disorder with an incidence of 1/10,000. The disease is characterized by involuntary choreic movements, psychiatric disorders, dementia, and death within 15-20 years. When the HD gene was cloned in 1993, it was discovered that the causative mutation was an expansion of a CAG trinucleotide repeat (1) and to date HD is one of 14 trinucleotide repeat diseases (2). The expanded CAG sequence in exon 1 of the HD gene, IT15, likewise encodes an expanded polyglutamine tract resulting in an aberrant huntingtin protein, which leads to neuronal specific death predominantly within the neostratum (3). Huntington is widely expressed in both the brain and nonneural tissues (4) and interestingly, N-terminal fragments of huntingtin, containing the elongated polyglutamine residues form aggregates, and can be visualized as cytoplasmic and nuclear inclusions (5).

Keywords

Polymerase Chain Reaction Buffer Plate Assembly Fluorescence Polymerase Chain Reaction Nonneural Tissue Autosomal Dominant Neurodegenerative Disorder 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Huntington’s Disease Collaborative Research Group (1993) A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell 72, 971–983.CrossRefGoogle Scholar
  2. 2.
    Cummings, C. J. and Zoghbi, H. Y. (2000) Fourteen and counting: unraveling trinucleotide repeat diseases. Hum. Mol. Gene. 9, 909–916.CrossRefGoogle Scholar
  3. 3.
    Reiner, A., Albin R. L., Anderson, K. D., et al. (1988) Differential loss of striatal projection neurons in Huntington disease. Proc. Natl. Acad. Sci. USA 85, 5733–5737.PubMedCrossRefGoogle Scholar
  4. 4.
    Davies, S. W., Turmaine, M., Cozens, B. A., et al. (1997) Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. Cell 90, 537–548.PubMedCrossRefGoogle Scholar
  5. 5.
    Strong, T. V., Tagle, D. A., Valdes, J. M., et al. (1993) Widespread expression of the human and rat Huntington’s disease gene in brain and nonneural tissues. Nat. Gene. 5, 259–265.CrossRefGoogle Scholar
  6. 6.
    Meissen, G. J., Myers, R. H., Mastromauro, C. A., et al. (1988) Predictive testing for Huntington’s disease with use of a linked DNA marker. N. Engl. J. Med. 318, 535–542.PubMedCrossRefGoogle Scholar
  7. 7.
    Duyao, M., Ambrose, C., Myers, R., et al. (1993) Trinucleotide repeat length instability and age of onset in Huntington’s disease. Nat. Gene. 4, 387–392.CrossRefGoogle Scholar
  8. 8.
    McNeil, S. M., Novelletto, A., Srinidhi, J., et al. (1997) Reduced penetrance of the Huntington’s disease mutation. Hum. Mol. Gene. 6, 775–779.CrossRefGoogle Scholar
  9. 9.
    Rubinsztein, D. C., Leggo, J., Coles, R., et al. (1996) Phenotypic characterization of individuals with 30-40 CAG repeats in the Huntington Disease (HD) gene reveals HD cases with 36 repeats and apparently normal elderly individuals with 36-39 repeats. Am. J. Hum. Gene. 59, 16–22.Google Scholar
  10. 10.
    Goldberg, Y. P., McMurray, C. T., Zeisler, J., et al. (1995) Increased instability of intermediate alleles in families with sporadic Huntington disease compared to similar sized intermediate alleles in the general population. Hum. Mol. Gene. 4, 1911–1918.CrossRefGoogle Scholar
  11. 11.
    Warner, J. P., Barron, L. H. Brock, D. J. H., et al. (1993) A new polymerase chain reaction (PCR) assay for the trinucleotide repeat that is unstable and expanded on Huntington’s disease chromosomes. Mol. Cell. Probes 7, 235–239.PubMedCrossRefGoogle Scholar
  12. 12.
    (1994) Guidelines for the molecular genetics predictive test in Huntington’s disease. International Huntington Association (IHA) and the World Federation of Neurology (WFN) Research Group on Huntington’s Chorea. Neurology 44, 1533–1536.Google Scholar

Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  • Cindy L. Vnencak-Jones
    • 1
  1. 1.Department of PathologyVanderbilt University Medical CenterNashville

Personalised recommendations