Direct detection of novel expanded trinucleotide repeats in the human genome

Abstract

Expansion of trinucleotide repeats can give rise to genetic disease. We have developed a technique, repeat expansion detection (RED), that can identify potentially pathological repeat expansion without prior knowledge of chromosomal location. Human genomic DNA is used as a template for a two–step cycling process that generates oligonucleotide multimers when expanded trinucleotide sequences are present at the level found in myotonic dystrophy and fragile–X patients. We have identified at least one new locus exhibiting trinucleotide expansion. Analysis of three families transmitting a long CTG repeat shows that the allele in these families corresponds to a locus on chromosome 18. RED constitutes a powerful tool to identify other diseases caused by this mechanism, particularly diseases associated with anticipation.

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References

  1. 1

    Webb, T.P., Bundey, S.E., Thake, A.I. & Todd, J. The Frequency of the Fragile X Chromosome Among Schoolchildren in Coventry. Am. J. med. Genet. 23, 573–580 (1986).

    CAS  Article  Google Scholar 

  2. 2

    Gustavson, K.H., Blomquist, H. & Holmgren, G. Prevalence of Fragile–X Syndrome in Mentally Retarded Children in a Swedish County. Am. J. med. Genet. 23, 581–588 (1986).

    CAS  Article  Google Scholar 

  3. 3

    Harper, P.S. Myotonic Dystrophy, 2nd edn (W. B. Saunders, London, 1989).

    Google Scholar 

  4. 4

    Barany, F. Genetic Disease Detection and DNA Amplification Using Cloned Thermostable Ligase. Proc. natn. Acad. Sci. U.S.A. 88, 189–193 (1991).

    CAS  Article  Google Scholar 

  5. 5

    Schalling, M. In Gene Expression in Neural Tissues. (ed. Conn, P. M.) 231–255 (Academic Press, New York, 1992).

    Google Scholar 

  6. 6

    Brook, J.D. et al. Molecular Basis of Myotonic Dystrophy: Expansion of a Trinucleotide (CTG) Repeat at the 3′ End of a Transcript Encoding a Protein Kinase Family Member. Cell 68, 799–808 (1992).

    CAS  Article  Google Scholar 

  7. 7

    Verkerk, A.J.M.H. et al. Identification of a Gene (FMR-1) Containing a CGG Repeat Coincident with a Breakpoint Cluster Region Exhibiting Length Variation in Fragile X Syndrome. Cell 65, 905–914 (1991).

    CAS  Article  Google Scholar 

  8. 8

    Hinds, H. et al. Tissue Specific Expression of FMR-1 Provides Evidence for a Functional Role in Fragile X Syndrome. Nature Genet. 3, 36–43 (1993).

    CAS  Article  Google Scholar 

  9. 9

    Fu, Y.-H. et al. An Unstable Triplet Repeat in a Gene Related to Myotonic Muscular Dystrophy. Science 255, 1256–1258 (1992).

    CAS  Article  Google Scholar 

  10. 10

    Mahadevan, M. et al. Myotonic Dystrophy Mutation: an Unstable CTG Repeat in the 3′ Untranslated Region of the Gene. Science 255, 1253–1255 (1992).

    CAS  Article  Google Scholar 

  11. 11

    Richards, R.I. & Sutherland, G.R. Heritable Unstable DNA Sequences. Nature Genet. 1, 7–9 (1992).

    CAS  Article  Google Scholar 

  12. 12

    La Spada, A.R., Wilson, E.M., Lubahn, D.B., Harding, A.E. & Fischbeck, K.H. Androgen Receptor Gene Mutations in X-linked Spinal and Bulbar Muscular Atrophy. Nature 352, 77–79 (1991).

    CAS  Article  Google Scholar 

  13. 13

    Biancalana, F., Serville, F., Pommier, J., Julien, J. & Mandel, J.L. Moderate Instability of the Trinucleotide Repeat in Spino Bulbar Muscular Atrophy. Human molec. Genet. 1, 255–258 (1992).

    CAS  Article  Google Scholar 

  14. 14

    The Huntington's disease Collaboration group. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosome. Cell 72, 1–20 (1993).

  15. 15

    Aslanidis, C. et al. Cloning of the Essential Myotonic Dystrophy Region: Mapping of the Putative Defect. Nature 355, 548–551 (1992).

    CAS  Article  Google Scholar 

  16. 16

    Buxton, J. et al. Detection of an Unstable Fragment of DNA Specific to Individuals with Myotonic Dystrophy. Nature 355, 547–548 (1992).

    CAS  Article  Google Scholar 

  17. 17

    Harley, H.G. et al. Expansion of an Unstable DNA Region and Phenotypic Variation in Myotonic Dystrophy. Nature 355, 545–546 (1992).

    CAS  Article  Google Scholar 

  18. 18

    Riggins, G.J. et al. Human Genes Containing Polymorphic Trinucleotide Repeats. Nature Genet. 2, 186–191 (1992).

    CAS  Article  Google Scholar 

  19. 19

    Beckmann, J.S. & Weber, J.L. Survey of Human and Rat Microsatellites. Genomics 12, 627–631 (1992).

    CAS  Article  Google Scholar 

  20. 20

    Sambrook, J., Fritsch, E. & Maniatis, T. Molecular Cloning: A Laboratory Manual 3rd edn (Cold Spring Harbor Press, New York, 1989).

    Google Scholar 

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Schalling, M., Hudson, T., Buetow, K. et al. Direct detection of novel expanded trinucleotide repeats in the human genome. Nat Genet 4, 135–139 (1993). https://doi.org/10.1038/ng0693-135

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