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Preimplantation single cell analyses of dystrophin gene deletions using whole genome amplification

Nature Genetics volume 6, pages 19–23 (1994)Cite this article

Abstract

Primer extension preamplification (PEP) increases the scope and capacity of single cell genetic diagnosis by generating sufficient template to perform multiple subsequent DNA analyses using the polymerase chain reaction. We report the simultaneous analysis of single cells at five commonly deleted dystrophin exons and at the ZFX/ZFY loci. Ninety three percent of PEP reactions with single amniocytes, chorionic villus cells and blastomeres were successful, and a blinded analysis of single lymphoblasts from affected males resulted in 93% diagnostic accuracy, demonstrating its applicability in preimplantation prevention of Duchenne muscular dystrophy. Transfer of unaffected male embryos and improved diagnostic reliability are achieved with the ability to perform replicate multilocus analyses from the same blastomere.

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References

  1. Handyside, A.H., Kontogianni, E.H., Hardy, K. & Winston, R.M.L. Pregnancies from btopsied human preimplantation embryos sexed by Y-specific DNA amplification. Nature 344, 768–770 (1990).

    Article  CAS  Google Scholar 

  2. Handyside, A.H., Lesko, J.G., Tarin, J., Winston, R. & Hughes, M.R. Birth of a normal girl following preimplantation diagnosis for Cystic Fibrosis. New Engl. J. Med. 327, 905–909 (1992).

    Article  CAS  Google Scholar 

  3. Grifo, J.A. et al. Pregnancy after embryo biopsy and coamplification of DNA from X and Y chromosomes. J. Am. med. Assoc. 268, 727–729 (1992).

    Article  CAS  Google Scholar 

  4. Chamberlain, J.S. & Caskey, C.T. Duchenne muscular dystrophy. in Current Neurology X (ed. Appel, S. H.). 65–103 (Year Book Medical Publishers, Chicago, (1990).

    Google Scholar 

  5. Witkowski, J. Dystrophin-related muscular dystrophies. J. child Neurol. 4, 251–271 (1990).

    Article  Google Scholar 

  6. Koenig, M. et al. Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell 50, 509–517 (1987).

    Article  CAS  Google Scholar 

  7. Chamberlain, J.S., Gibbs, R.A., Ranier, J.E. & Caskey, C.T. Detection of gene deletions using multiplex polymerase chain reactions in Methods In Molecular Biology, IX: Protocols In Human Molecular Genetics (ed. Mathew, C.) 299–312 (Humana, Clifton, New Jersey, (1991).

    Chapter  Google Scholar 

  8. Beggs, A.H., Koenig, M., Boyce, F.M. & Kunkel, L.M. Detection of 98% of DMD/BMD gene deletions by polymerase chain reaction. Hum. Genet. 86, 45–48 (1990).

    Article  CAS  Google Scholar 

  9. Multicenter. Diagnosis of Duchenne and Becker muscular dystrophies by polymerase chain reaction. J. Am. med. Assoc. 267, 2609–2615 (1992).

  10. Griffin, D.K., Wilton, L.J., Handyside, A.H., Winston, R.M.L. & Delhanty, J.D.A. Dual fluorescent in-situ hybridization for simultaneous detection of X and Y chromosome-specific probes for the sexing of human preimplantation embryonic nuclei. Hum. Genet. 89, 18–22 (1992).

    Article  CAS  Google Scholar 

  11. Chong, S.S., Kristjansson, K., Cota, J., Handyside, A.H. & Hughes, M.R. Preimplantation prevention of X-linked disease: reliable and rapid sex determination of single human cells by restriction analysis of simultaneously amplified ZFX and ZFY sequences. Hum. molec. Genet. 2, 1187–1192 (1993).

    Article  CAS  Google Scholar 

  12. Zhang, L. et al. Whole genome amplification from a single cell: Implications for genetic analysts. Proc. natn. Acad. Sci. U.S.A. 89, 5847–5851 (1992).

    Article  CAS  Google Scholar 

  13. Hubert, R., Weber, J.L., Schmitt, K., Zhang, L. & Arnheim, N. A new source of polymorphic DNA markers for sperm typing: analysis of microsatellite repeats in single cells. Am. J. hum. Genet. 51, 985–991 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Navidi, W., Arnheim, N. & Waterman, M.S. A multiple-tubes approach for accurate genotyping of very small DNA samples by using PCR: statistical considerations. Am. J. hum. Genet. 50, 347–359 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Navidi, W. & Arnheim, N. Using PCR in preimplantation genetic disease diagnosis. Hum. Reprod. 6, 836–849 (1991).

    Article  CAS  Google Scholar 

  16. Kontogianni, E.H., Hardy, K. & Handyside, A.H. Co-amplification of X-and Y-specific sequences for sexing preimplantation human embryos. in Preimplantation Genetics (eds Verlinsky, Y. & Kuliev, A.) 139–145 (Plenum, New York, (1991).

    Chapter  Google Scholar 

  17. Pickering, S.J., McConnell, J.M., Johnson, M.H. & Braude, P.R. Reliability of detection by polymerase chain reaction of the sickle cell-containing region of the β-globin gene in single human blastomeres. Hum. Reprod. 7, 630–636 (1992).

    Article  CAS  Google Scholar 

  18. Cui, X. et al. Single sperm typing: determination of genetic distance between the Gγ-globin and parathyroid hormone loci. Proc. natn. Acad. Sci. U.S.A. 86, 9389–9393 (1989).

    Article  CAS  Google Scholar 

  19. Li, A. et al. Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature 335, 414–419 (1988).

    Article  CAS  Google Scholar 

  20. Boehnke, M., Arnheim, N., Li, H. & Collins, F.S. Fine-structure genetic mapping of human chromosomes using the polymerase chain reaction on single sperm: experimental design considerations. Am. J. hum. Genet. 45, 21–32 (1989).

    CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas, 77030, USA

    Kristleifur Kristjansson, Samuel S. Chong, Ignatia B. Van den Veyver, Stephanie Subramanian & Mark R. Hughes

  2. Department of Obstetrics and Gynecology, Baylor College of Medicine, Houston, Texas, 77030, USA

    Michael C. Snabes

Authors
  1. Kristleifur Kristjansson
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  2. Samuel S. Chong
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  3. Ignatia B. Van den Veyver
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  4. Stephanie Subramanian
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  5. Michael C. Snabes
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  6. Mark R. Hughes
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Kristjansson, K., Chong, S., Van den Veyver, I. et al. Preimplantation single cell analyses of dystrophin gene deletions using whole genome amplification. Nat Genet 6, 19–23 (1994). https://doi.org/10.1038/ng0194-19

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