Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Jun;87(12):4481–4484. doi: 10.1073/pnas.87.12.4481

Rapid preimplantation detection of mutant (shiverer) and normal alleles of the mouse myelin basic protein gene allowing selective implantation and birth of live young.

C M Gomez 1, A L Muggleton-Harris 1, D G Whittingham 1, L E Hood 1, C Readhead 1
PMCID: PMC54139  PMID: 1693773

Abstract

As a model for the detection of human genetic disease in preimplantation embryos, we describe a method in which trophectoderm biopsy samples from viable mouse blastocysts are simultaneously analyzed for the presence of a normal or mutant allele of the myelin basic protein gene by the polymerase chain reaction. The biopsied embryos are kept in culture during analysis of biopsied material and later reintroduced to a foster mother. Prenatal diagnosis can be completed in less than 7 hr. The identity of either amplification product was proved conclusively by direct sequence analysis of amplified products. Ninety-six percent of recovered blastocysts survived biopsy, as judged by re-formation of a blastocyst cavity in culture. Fifty-nine percent of the biopsied embryos established pregnancy by day 6.5, compared to 88% of unmanipulated controls. This approach can be applied to preimplantation diagnosis of human genetic diseases by using extraembryonic cells from blastocysts obtained after in vitro fertilization or uterine lavage. It will make possible the elimination of a mutant allele from a family in a single generation.

Full text

PDF
4481

Images in this article

4482Image
on p.4482
4482Image
on p.4482
4482Image
on p.4482
4483Image
on p.4483

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Handyside A. H., Pattinson J. K., Penketh R. J., Delhanty J. D., Winston R. M., Tuddenham E. G. Biopsy of human preimplantation embryos and sexing by DNA amplification. Lancet. 1989 Feb 18;1(8634):347–349. doi: 10.1016/s0140-6736(89)91723-6. [DOI] [PubMed] [Google Scholar]
  2. Holding C., Monk M. Diagnosis of beta-thalassaemia by DNA amplification in single blastomeres from mouse preimplantation embryos. Lancet. 1989 Sep 2;2(8662):532–535. doi: 10.1016/s0140-6736(89)90655-7. [DOI] [PubMed] [Google Scholar]
  3. Kretz K. A., Carson G. S., O'Brien J. S. Direct sequencing from low-melt agarose with Sequenase. Nucleic Acids Res. 1989 Jul 25;17(14):5864–5864. doi: 10.1093/nar/17.14.5864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  5. Landegren U., Kaiser R., Caskey C. T., Hood L. DNA diagnostics--molecular techniques and automation. Science. 1988 Oct 14;242(4876):229–237. doi: 10.1126/science.3051381. [DOI] [PubMed] [Google Scholar]
  6. Landegren U., Kaiser R., Sanders J., Hood L. A ligase-mediated gene detection technique. Science. 1988 Aug 26;241(4869):1077–1080. doi: 10.1126/science.3413476. [DOI] [PubMed] [Google Scholar]
  7. Li H. H., Gyllensten U. B., Cui X. F., Saiki R. K., Erlich H. A., Arnheim N. Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature. 1988 Sep 29;335(6189):414–417. doi: 10.1038/335414a0. [DOI] [PubMed] [Google Scholar]
  8. Molineaux S. M., Engh H., de Ferra F., Hudson L., Lazzarini R. A. Recombination within the myelin basic protein gene created the dysmyelinating shiverer mouse mutation. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7542–7546. doi: 10.1073/pnas.83.19.7542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Monk M., Muggleton-Harris A. L., Rawlings E., Whittingham D. G. Pre-implantation diagnosis of HPRT-deficient male and carrier female mouse embryos by trophectoderm biopsy. Hum Reprod. 1988 Apr;3(3):377–381. doi: 10.1093/oxfordjournals.humrep.a136711. [DOI] [PubMed] [Google Scholar]
  10. Quinn P., Barros C., Whittingham D. G. Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa. J Reprod Fertil. 1982 Sep;66(1):161–168. doi: 10.1530/jrf.0.0660161. [DOI] [PubMed] [Google Scholar]
  11. Roach A., Boylan K., Horvath S., Prusiner S. B., Hood L. E. Characterization of cloned cDNA representing rat myelin basic protein: absence of expression in brain of shiverer mutant mice. Cell. 1983 Oct;34(3):799–806. doi: 10.1016/0092-8674(83)90536-6. [DOI] [PubMed] [Google Scholar]
  12. Takahashi N., Roach A., Teplow D. B., Prusiner S. B., Hood L. Cloning and characterization of the myelin basic protein gene from mouse: one gene can encode both 14 kd and 18.5 kd MBPs by alternate use of exons. Cell. 1985 Aug;42(1):139–148. doi: 10.1016/s0092-8674(85)80109-4. [DOI] [PubMed] [Google Scholar]
  13. Whittingham D. G. Culture of mouse ova. J Reprod Fertil Suppl. 1971 Jun;14:7–21. [PubMed] [Google Scholar]
  14. de Ferra F., Engh H., Hudson L., Kamholz J., Puckett C., Molineaux S., Lazzarini R. A. Alternative splicing accounts for the four forms of myelin basic protein. Cell. 1985 Dec;43(3 Pt 2):721–727. doi: 10.1016/0092-8674(85)90245-4. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES