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.

  • Review Article
  • Published:

Modifying the Mouse: Design and Desire

Abstract

Genetic modification of endogenous genes in mice has become possible by applying gene targeting techniques to embryonic stem (ES) cells and using specific clones of cells to generate mice. Despite the experimental opportunities offered by the creation of organisms with specific genetic changes, there are considerable technical obstacles which can confound the routine implementation of this technology. This review addresses some recent advances in the ability to construct mice with a variety of genetic modifications. These include an increased understanding of the basic cell biology and in vitro growth characteristics of ES cells, which has facilitated germ line transmission of manipulated clones on a routine basis. The techniques that are used to isolate “targeted” clones of ES cells have been summarized, and the current status of strategies which have been successfully used to make very specific modifications of the genome are discussed.

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

Access options

Buy this article

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

Similar content being viewed by others

References

  1. Palmiter, R.D. and Brinster, R.L. 1986. Germ line transformation of mice. Ann. Rev. Genet. 20: 465–499.

    Article  CAS  Google Scholar 

  2. Evans, M.J. and Kaufman, M.H. 1981. Establishment in culture of pluripotential cells from mouse embryos. Nature 292: 154–156.

    Article  CAS  Google Scholar 

  3. Bradley, A., Evans, M.J., Kaufman, M.H. and Robertson, E.J. 1984. Formation of germ line chimaeras from embryo-derived teratocarcinoma cell lines. Nature 309: 255–256.

    Article  CAS  Google Scholar 

  4. Thomas, K.R. and Capecchi, M.R. 1987. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51: 503–512.

    Article  CAS  Google Scholar 

  5. Kuehn, M.R., Bradley, A., Robertson, E.J. and Evans, M.D. 1987. A potential animal model for Lesch-Nyhan syndrome through introductions of hprt mutations into mice. Nature 326: 295–298.

    Article  CAS  Google Scholar 

  6. Schwartzberg, P.I., Goff, S.P. and Robertson, E.J. 1989. Germ-line transmission of a c-abl mutation produced by targeted disruption in ES cells. Science 246: 799–804.

    Article  CAS  Google Scholar 

  7. Robertson, E.J., Bradley, A., Kuehn, M. and Evans, M.J. 1986. Germ line transmission of genes introduced into cultured pluripotential cells by a retroviral vector. Nature 323: 445–448.

    Article  CAS  Google Scholar 

  8. Bradley, A. 1990. Embryonic stem cells: Proliferation and differentiation. Current Opinion in Cell Biology 2: 1013–1017.

    Article  CAS  Google Scholar 

  9. Robertson, E.J. 1987. Embryo-derived stem cells lines, 71–112. In: Teratocarcinomas and Embryonic Stem Cells: A Practical Approach. E.J. Robertson (Ed.). IRL Press, Oxford, UK. .

    Google Scholar 

  10. Smith, A.G. and Hooper, M.L. 1987. Buffalo rat liver cells produce a diffusible activity which inhibits the differentiation of embryonal carcinoma and embryonic stem cells. Dev. Biol. 121: 1–9.

    Article  CAS  Google Scholar 

  11. Williams, R.L., Hilton, D.J., Pease, S., Willison, T.A., Stewart, C.L., Gearing, D.P., Wagner, E.F., Metcalf, D., Nicola, N.A. and Gough, N.M. 1988. Myeloid leukemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336: 684–686.

    Article  CAS  Google Scholar 

  12. Pease, S. and Williams, R.L. 1990. Formation of germ line chimaeras from embryonic stem cells maintained with recombinant leukemia inhibitory factor. Exp. Cell Res. 190: 209–211.

    Article  CAS  Google Scholar 

  13. Zijlstra, M., Li, E., Sajjadi, F., Subramani, S. and Jaenisch, R. 1989. Germ-line transmission of a disrupted β2-microglobulin gene produced by homologous recombination in embryonic stem cells. Nature 342: 435–438.

    Article  CAS  Google Scholar 

  14. McMahon, A.P. and Bradley, A. 1990. The Wnt-1 (Int-1) proto-oncogene is required for the development of a large region of the mouse brain. Cell 62: 1073–1085.

    Article  CAS  Google Scholar 

  15. Stanton, B.R., Reid, S.W. and Parada, L.F. 1990. Germ line transmission of an inactive N-myc allele generated by homologous recombination in mouse embryonic stem cells. Mol. Cell Biol. 10: 6755–6758.

    Article  CAS  Google Scholar 

  16. Tybulewicz, V.L.J., Crawford, C.E., Jackson, P.K., Bronson, R.T. and Mulligan, R.C. 1991. Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene. Cell 65: 1153–1164.

    Article  CAS  Google Scholar 

  17. Soriano, P., Montgomery, C., Geske, R. and Bradley, A. 1991. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell 64: 693–702.

    Article  CAS  Google Scholar 

  18. Thomas, K.R. and Capecchi, M.R. 1990. Targeted disruption of the murine int-1 proto-onco gene resulting in severe abnormalities in the midbrain and cerebellar development. Nature 346: 847–850.

    Article  CAS  Google Scholar 

  19. teRiele, H., Maandag, E.R., Clarke, A., Hooper, M. and Berns, A. 1990. Consecutive inactivation of both alleles of the pim-1 proto-oncogene by homologous recombination in embryonic stem cells. Nature 348: 649–651.

    Article  CAS  Google Scholar 

  20. Friedrich, G. and Soriano, P. 1991. Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes & Dev. 5: 1513–1523.

    Article  CAS  Google Scholar 

  21. DeChira, T.M., Efstatiadis, A. and Robertson, E.J. 1990. A growth deficiency phenotype in heterozygous mice carrying an insulin like growth factor II gene disrupted by targeting. Nature 345: 78–80.

    Article  Google Scholar 

  22. Chiska, O. and Capecchi, M.R. 1991. Regionally restricted developmental defects resulting from targeted disruption of the mouse homeobox gene Hox 1.5. Nature 350: 473–479.

    Article  Google Scholar 

  23. Thompson, S., Clarke, A.R., Pow, A., Hooper, M.L. and Melton, D.W. 1989. Germ-line transmission and expression of a corrected HPRT gene produced by gene targeting in embryonic stem cells. Cell 56: 313–321.

    Article  CAS  Google Scholar 

  24. Joyner, A.L., Auerbach, B.A., Davis, C.A., Herrup, K. and Rossant, J. 1991. Subtle cerebellar phenotype in mice homozygous for a targeted deletion of the En2 homedomain. Science 251: 1239–1243.

    Article  CAS  Google Scholar 

  25. Hasty, P., Ramírez-Solis, R., Krumlauf, R. and Bradley, A. 1991. Introduction of a subtle mutation into the Hax-2.6 locus in embryonic stem cells. Nature 350: 243–246.

    Article  CAS  Google Scholar 

  26. Smithies, O., Gregg, R.G., Boggs, S.S., Koralewski, M.A. and Kucherlapati, R.S. 1985. Insertion of DNA sequences into the human chromosomal β-globin locus by homologous recombination. Nature 317: 230–234.

    Article  CAS  Google Scholar 

  27. Hasty, P., Rivera-Pérez, J. and Bradley, A. 1991. The length of homology required for gene targeting in embryonic stem cells. Mol. Cell Biol. 11: 5586–5591.

    Article  CAS  Google Scholar 

  28. Hasty, P., Rivera-Pérez, J., Chang, C. and Bradley, A. 1991. Target frequency and integration pattern for insertion and replacement vectors in embryonic stem cells. Mol. Cell. Bio. 11: 4509–4517.

    Article  CAS  Google Scholar 

  29. Schwartzberg, P.L., Robertson, E.J. and Goff, S.P. 1990. Targeted disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein. Proc. Natl. Acad. Sci. U.S.A. 87: 3210–3214.

    Article  CAS  Google Scholar 

  30. Charron, J., Malynn, B.A., Robertson, E.J., Goff, S.P. and Alt, F.W. 1990. High-frequency disruption of the N-myc gene in embryonic stem and pre-B cell lines by homologous recombination. Mol. Cell Biol. 10: 1799–8084.

    Article  CAS  Google Scholar 

  31. Donehower, L.A., Harvey, M., Slagle, B., McArthur, M.J., Montgomery, C.A., Butel, J.S. and Bradley, A. 1992. Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature 356: 215–221.

    Article  CAS  Google Scholar 

  32. Mansour, S.L., Thomas, K.R. and Capecchi, M.R. 1988. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 336: 348–352.

    Article  CAS  Google Scholar 

  33. Joyner, A.L., Skarnes, W.C. and Rossant, J. 1989. Production of a mutation in the mouse En2 gene by homologous recombination in embryonic stem cells. Nature 338: 153–156.

    Article  CAS  Google Scholar 

  34. Ramirez-Solis, R., Rivera-Perez, J., Wallace, J.D., Wims, M., Zheng, H. and Bradley, A. 1992. Genomic DNA microextraction: a method to screen numerous samples. Analytical Biochemistry 201: 331–335.

    Article  CAS  Google Scholar 

  35. Mansour, S.L., Thomas, K.R., Deng, C.X. and Capecchi, M.R. 1990. Introduction of a lacZ reporter gene into the mouse int-2 locus by homologous recombination. Proc. Natl. Acad. Sci. U.S.A. 87: 7688–7692.

    Article  CAS  Google Scholar 

  36. Le-Mouellic, H., Lallemand, Y. and Brulet, P. 1990. Targeted gene replacement of the homeobox gene Hox-3.1by the Escherichia coli lacZ in mouse chimeric embryos. Proc. Natl. Acad. Sci. U.S.A. 87: 4712–4716.

    Article  CAS  Google Scholar 

  37. Zimmer, A. and Gruss, P. 1989. Production of chimaeric mice containing embryonic stem (ES) cells carrying a homeobox Hox1.1 allele mutated by homologous recombination. Nature 338: 150–153.

    Article  CAS  Google Scholar 

  38. Davis, A.C. and Bradley, A. 1992. Co-electroporation as a method for introducing small mutations into embryonic stem cells. Mol. Cell Biol. In press

  39. Shesely, E.G., Kim, H.S., Shehee, W.R., Papayannopoulou, T., Smithies, O. and Popovich, B.W. 1991. Correction of a human beta S-globin gene by gene targeting. Proc. Natl. Acad. Sci. U.S.A. 88: 4294–4298.

    Article  CAS  Google Scholar 

  40. Schnieke, A., Harbers, K. and Jaenisch, R. 1983. Embryonic lethal mutation in mice induced by retrovirus insertion into the α1(l) collagen gene. Nature 304: 315–320.

    Article  CAS  Google Scholar 

  41. Nusse, R. 1986. The activation of cellular oncogenes by retroviral insertion. Trends Genet 2: 244–247.

    Article  CAS  Google Scholar 

  42. Valancius, V. and Smithies, O. 1991. Testing an “In-Out” targeting procedure for making subtle genomic modifications in mouse embryonic stem cells. Mol. Cell Biol. 11: 1402–1408.

    Article  CAS  Google Scholar 

  43. Bradley, A., Ramirez-Solis, R., Zheng, H., Hasty, P. and Davis, A.C. 1992. Genetic manipulation of the mouse via gene targeting in embryonic stem cells. In: Ciba Foundation-Post Implantation Development of the Mouse. In press.

    Google Scholar 

  44. Grosveld, F., Blom van Assendelft, G., Greaves, D.R. and Kollias, G. 1987. Position-independent, high level expression of the human β-globin gene in transgenic mice. Cell 51: 975–985.

    Article  CAS  Google Scholar 

  45. Pavan, W.J., Hieter, P. and Reeves, R.H. 1990. Modification and transfer into an embryonal carcinoma cell line of a 360-kilobase human derived artificial chromosomes. Mol. Cell. Bio. 10: 4163–4169.

    Article  CAS  Google Scholar 

  46. Notarianni, E., Laurie, S., Moor, R. and Evans, M.J. 1990. Maintenance and differentiation in culture of pluripotential embryonic cell lines from pig blastocysts. J. Reprod. Fert. Suppl. 41: 51–56.

    CAS  Google Scholar 

  47. Piedrahita, J.A., Anderson, G.B. and BonDurant, R.H. 1990. On the isolation of embryonic stem cells: Comparative behaviour of murine, porcine and ovine embryos. Theriogenology 34: 865–878.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bradley, A., Hasty, P., Davis, A. et al. Modifying the Mouse: Design and Desire. Nat Biotechnol 10, 534–539 (1992). https://doi.org/10.1038/nbt0592-534

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0592-534

This article is cited by

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