Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Technical Report
  • Published:

Efficient metaphase II transgenesis with different transgene archetypes

Nature Biotechnology volume 19, pages 1071–1073 (2001)Cite this article

Abstract

Mammalian genome characterization and biotechnology each require the mobilization of large DNA segments to produce transgenic animals. We recently showed that mouse metaphase II (mII) oocytes could efficiently promote transgenesis (mII transgenesis) when coinjected with sperm and small (<5 kilobases) ubiquitously expressed transgenes (tgs)1. We have extended this work and now report that mII transgenesis can readily be applied to a range of larger tgs (11.9–170 kilobases), including bacterial and mammalian artificial chromosome (BAC and MAC) constructs. The efficiency of large-construct mII transgenesis was at least as high as that with small constructs; 11–47% of offspring carried the large tgs. More than 95% of these transgenic founders transmitted the tg to offspring. These data demonstrate the ability of mII transgenesis to deliver large tgs efficiently.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Structure of pTATG and FISH analysis of a pTATG transgenic integrant.

Similar content being viewed by others

References

  1. Perry, A.C.F. et al. Mammalian transgenesis by intracytoplasmic sperm injection. Science 284, 1180–1183 (1999).

    Article  CAS  Google Scholar 

  2. Ebersole, T.A. et al. Mammalian artificial chromosome formation from circular alphoid input DNA does not require telomere repeats. Hum. Mol. Genet. 9, 1623–1631 (2000).

    Article  CAS  Google Scholar 

  3. Kuretake, S., Kimura, Y., Hoshi, K. & Yanagimachi, R. Fertilization and development of mouse oocytes injected with isolated sperm heads. Biol. Reprod. 55, 789–795 (1996).

    Article  CAS  Google Scholar 

  4. Wakayama, T., Whittingham, D.G. & Yanagimachi, R. Production of normal offspring from mouse oocytes injected with spermatozoa cryopreserved with or without cryoprotection. J. Fert. Reprod. 112, 11 (1998).

    Article  CAS  Google Scholar 

  5. Zhou, B.-B. & Elledge, S.J. The DNA damage response: putting checkpoints in perspective. Nature 408, 433–439 (2000).

    Article  CAS  Google Scholar 

  6. Perry, A.C.F. Hijacking oocyte DNA repair machinery in transgenesis? Mol. Reprod. Dev. 56, 319–324 (2000).

    Article  CAS  Google Scholar 

  7. Brinster, R.L., Chen, H.Y., Trumbauer, M.E., Yagle, M.K. & Palmiter, R.D. Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc. Natl. Acad. Sci. USA 82, 4438–4442 (1985).

    Article  CAS  Google Scholar 

  8. Takada, T. et al. Selective production of transgenic mice using green fluorescent protein as a marker. Nat. Biotechnol. 15, 458–461 (1997).

    Article  CAS  Google Scholar 

  9. Giraldo, P. & Montoliu, L. Size matters: use of YACs, BACs and PACs in transgenic animals. Transgenic Res. 10, 83–103 (2001).

    Article  CAS  Google Scholar 

  10. Yang, X.W., Model, P. & Heintz, N. Homologous recombination based modification in Escherichia coli and germline transmission in transgenic mice of a bacterial artificial chromosome. Nat. Biotechnol. 15, 859–865 (1997).

    Article  CAS  Google Scholar 

  11. Kimura, Y. & Yanagimachi, R. Intracytoplasmic sperm injection in the mouse. Biol. Reprod. 52, 709–720 (1995).

    Article  CAS  Google Scholar 

  12. Chatot, C.L., Lewis, J.L., Torres, I. & Ziomek, C.A. Development of 1-cell embryos from different strains of mice in CZB medium. Biol. Reprod. 42, 432–440 (1990).

    Article  CAS  Google Scholar 

  13. Fantes, J.A. et al. Submicroscopic deletions at the WAGR locus revealed by nonradioactive in situ hybridization. Am. J. Hum. Genet. 51, 1286–1294 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported in part by a generous contribution from The Rockefeller University. J.I. de las H. and H.J.C. are supported by the Medical Research Council (UK). We are grateful for assistance from I. Rodriguez, M. Fernandes, C. Zheng, and R. Nazar.

Author information

Author notes

  1. Anthony C.F. Perry & Teruhiko Wakayama

    Present address: Advanced Cell Technology, One Innovation Drive, Worcester, MA, 01605, USA

Authors and Affiliations

  1. The Rockefeller University, 1230 York Avenue, New York, 10021, NY, USA

    Anthony C.F. Perry, Andrea Rothman, Paul Feinstein, Peter Mombaerts & Teruhiko Wakayama

  2. Chromosome Biology Section, MRC Human Molecular Genetics Unit, Edinburgh, EH4 2XU, UK

    Jose I. de las Heras & Howard J. Cooke

Authors
  1. Anthony C.F. Perry
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrea Rothman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jose I. de las Heras
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paul Feinstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peter Mombaerts
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Howard J. Cooke
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Teruhiko Wakayama
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anthony C.F. Perry.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Perry, A., Rothman, A., de las Heras, J. et al. Efficient metaphase II transgenesis with different transgene archetypes. Nat Biotechnol 19, 1071–1073 (2001). https://doi.org/10.1038/nbt1101-1071

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1101-1071

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing