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Repeat Expansion Detection (RED) and the RED Cloning Strategy

  • Qiu-Ping Yuan
  • Kerstin Lindblad-Toh
  • Martin Schalling
Part of the Methods in Molecular Biology™ book series (MIMB, volume 217)

Abstract

Trinucleotide repeat sequences are present at approx 30,000-40,000 loci in the human genome (1). The majority of these repeats are below 35 copies and are stably transmitted. However, unstable trinucleotide repeat expansions at some loci have been found to be the causal mutation for nearly 20 genetic neurodegenerative disorders in human (2, 3, 4). Most of these disorders occur at repeat lengths above 35 copies, with a tendency towards further expansion upon successive transmissions. An inverse correlation between the repeat length and disease severity/earlier age of onset, known as anticipation, has been observed in most of the families transmitting such types of diseases, suggesting that the length change of the repeats may play a role in the manifestation of anticipation. Only three motifs, CAG/CTG, CGG/CCG, and GAA/TTC, of the 10 possible trinucleotide repeat permutations have so far been associated with human disease. It remains possible that other disease phenotypes are caused by expansions of any repeat motif at any repeat containing locus. We have established a repeat detection and gene-isolation system, which allows identification of a repeat-containing gene within a couple of months.

Keywords

Sodium Dodecyl Sulfate Repeat Length Trinucleotide Repeat Cloning Procedure Reaction Failure 
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.
    Lander, E. S., Linton, L. M., Birren, B., Nusbaum, C., Zody, M. C., Baldwin, J., et al. (2001) Initial sequencing and analysis of the human genome. Nature 409, 860–921.PubMedCrossRefGoogle Scholar
  2. 2.
    Warren, S. T. and Wells, R. D. (1998) Genetic Instabilities and Hereditary Neurological Diseases. Academic Press, San Diego, CA, USA.Google Scholar
  3. 3.
    Bowater, R. P. and Wells, R. D. (2000) The intrinsically unstable life of DNA triplet repeats associated with human hereditary disorders. Prog. Nucleic. Acid. Res. Mol. Biol. 66, 159–202.CrossRefGoogle Scholar
  4. 4.
    Zoghbi, H. Y. and Orr, H. T. (2000) Glutamine repeats and neurodegeneration. Annu. Rev. Neurosci. 23, 217–247.PubMedCrossRefGoogle Scholar
  5. 5.
    Schalling, M., Hudson, T. J., Buetow, K. H., and Housman, D. E. (1993) Direct detection of novel expanded trinucleotide repeats in the human genome. Nat. Genet. 4, 135–139.PubMedCrossRefGoogle Scholar
  6. 6.
    Zander, C., Thelaus, J., Lindblad, K., Karlsson, M., Sjoberg, K., and Schalling, M. (1998) Multivariate analysis of factors influencing repeat expansion detection. Genome Res. 8, 1085–1094.PubMedGoogle Scholar
  7. 7.
    Lindblad, K., Nylander, P. O., De bruyn, A., Sourey, D., Zander, C., Engstrom, C., et al. (1995) Detection of expanded CAG repeats in bipolar affective disorder using the repeat expansion detection (RED) method. Neurobiol. Dis. 2, 55–62.PubMedCrossRefGoogle Scholar
  8. 8.
    Lindblad, K., Savontaus, M. L., Stevanin, G., Holmberg, M., Digre, K., Zander, C., et al. (1996) An expanded CAG repeat sequence in spinocerebellar ataxia type 7. Genome Res. 6, 965–971.PubMedCrossRefGoogle Scholar
  9. 9.
    Lindblad, K., Lunkes, A., Maciel, P., Stevanin, G., Zander, C., Klockgether, T., et al. (1996) Mutation detection in Machado-Joseph disease using repeat expansion detection. Mol. Med. 2, 77–85.PubMedGoogle Scholar
  10. 10.
    Holmes, S. E., O’Hearn, E. E., McInnis, M.G., Gorelick-Feldman, D. A., Kleiderlein, J. J., Callahan, C., et al. (1999) Expansion of a novel CAG trinucleotide repeat in the 5’ region of PPP2R2B is associated with SCA12. Nat. Genet. 23, 391–392.PubMedCrossRefGoogle Scholar
  11. 11.
    Wells, R. D. (1996) Molecular basis of genetic instability of triplet repeats. J. Biol. Chem. 271, 2875–2878.PubMedGoogle Scholar
  12. 12.
    Iyer, R. R. and Wells, R. D. (1999) Expansion and deletion of triplet repeat sequences in Escherichia coli occur on the leading strand of DNA. J. Biol. Chem. 274, 3865–3877.PubMedCrossRefGoogle Scholar
  13. 13.
    Yuan, Q. P., Lindblad, K., Zander, C., Burgess, C., Durr, A., and Schalling, M. (2001) A cloning strategy for the identification of novel genes containing trinucleotide repeat expansion. Int. J. Med. Genet. 8, 427–431Google Scholar

Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  • Qiu-Ping Yuan
    • 1
  • Kerstin Lindblad-Toh
    • 1
  • Martin Schalling
    • 1
  1. 1.Department of Molecular MedicineKarolinska InstituteStockholmSweden

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