Skip to main content
NCBI 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
. 1989 Apr;86(8):2698–2702. doi: 10.1073/pnas.86.8.2698

Thymidine kinase obliteration: creation of transgenic mice with controlled immune deficiency.

R A Heyman 1, E Borrelli 1, J Lesley 1, D Anderson 1, D D Richman 1, S M Baird 1, R Hyman 1, R M Evans 1
PMCID: PMC286985  PMID: 2539597

Abstract

The cell-specific expression of herpes simplex virus 1 thymidine kinase (HSV-1-tk) has provided a simple and highly efficient technique to achieve conditional ablation of targeted cell types in transgenic mice. The ablation is induced by treating transgenic animals expressing HSV-1-tk with the antiherpetic drug ganciclovir. In lymphoid tissues of mice expressing HSV-1-tk from an immunoglobulin promoter, administration of ganciclovir leads to massive destruction of B- and T-cell lineages. Tissues not expressing HSV-1-tk are insensitive to drug treatment. After depletion of greater than 99% of total thymocytes, a number of progenitor cells remain that are able to repopulate all T-cell lineages within 7 days. The ability to control and direct ablation allows for creation of conditional mutant phenotypes at precise periods of development. This technique also provides a potential means to enrich stem cell populations as well as permitting the creation of animal models for particular pathological conditions.

Full text

PDF
2698

Images in this article

2700Image
on p.2700

Selected References

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

  1. Banerji J., Olson L., Schaffner W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell. 1983 Jul;33(3):729–740. doi: 10.1016/0092-8674(83)90015-6. [DOI] [PubMed] [Google Scholar]
  2. Behringer R. R., Mathews L. S., Palmiter R. D., Brinster R. L. Dwarf mice produced by genetic ablation of growth hormone-expressing cells. Genes Dev. 1988 Apr;2(4):453–461. doi: 10.1101/gad.2.4.453. [DOI] [PubMed] [Google Scholar]
  3. Bluestone J. A., Pardoll D., Sharrow S. O., Fowlkes B. J. Characterization of murine thymocytes with CD3-associated T-cell receptor structures. Nature. 1987 Mar 5;326(6108):82–84. doi: 10.1038/326082a0. [DOI] [PubMed] [Google Scholar]
  4. Borrelli E., Heyman R., Hsi M., Evans R. M. Targeting of an inducible toxic phenotype in animal cells. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7572–7576. doi: 10.1073/pnas.85.20.7572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breitman M. L., Clapoff S., Rossant J., Tsui L. C., Glode L. M., Maxwell I. H., Bernstein A. Genetic ablation: targeted expression of a toxin gene causes microphthalmia in transgenic mice. Science. 1987 Dec 11;238(4833):1563–1565. doi: 10.1126/science.3685993. [DOI] [PubMed] [Google Scholar]
  6. Crispe I. N., Moore M. W., Husmann L. A., Smith L., Bevan M. J., Shimonkevitz R. P. Differentiation potential of subsets of CD4-8- thymocytes. Nature. 1987 Sep 24;329(6137):336–339. doi: 10.1038/329336a0. [DOI] [PubMed] [Google Scholar]
  7. Elion G. B., Furman P. A., Fyfe J. A., de Miranda P., Beauchamp L., Schaeffer H. J. Selectivity of action of an antiherpetic agent, 9-(2-hydroxyethoxymethyl) guanine. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5716–5720. doi: 10.1073/pnas.74.12.5716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ewing T., Egerton M., Wilson A., Scollay R., Shortman K. Subpopulations of CD4- CD8- murine thymocytes: differences in proliferation rate in vivo and proliferative responses in vitro. Eur J Immunol. 1988 Feb;18(2):261–268. doi: 10.1002/eji.1830180213. [DOI] [PubMed] [Google Scholar]
  9. Fowlkes B. J., Edison L., Mathieson B. J., Chused T. M. Early T lymphocytes. Differentiation in vivo of adult intrathymic precursor cells. J Exp Med. 1985 Sep 1;162(3):802–822. doi: 10.1084/jem.162.3.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fowlkes B. J., Kruisbeek A. M., Ton-That H., Weston M. A., Coligan J. E., Schwartz R. H., Pardoll D. M. A novel population of T-cell receptor alpha beta-bearing thymocytes which predominantly expresses a single V beta gene family. Nature. 1987 Sep 17;329(6136):251–254. doi: 10.1038/329251a0. [DOI] [PubMed] [Google Scholar]
  11. Freitas A. A., Rocha B., Coutinho A. A. Lymphocyte population kinetics in the mouse. Immunol Rev. 1986 Jun;91:5–37. doi: 10.1111/j.1600-065x.1986.tb01482.x. [DOI] [PubMed] [Google Scholar]
  12. Furman P. A., McGuirt P. V., Keller P. M., Fyfe J. A., Elion G. B. Inhibition by acyclovir of cell growth and DNA synthesis of cells biochemically transformed with herpesvirus genetic information. Virology. 1980 Apr 30;102(2):420–430. doi: 10.1016/0042-6822(80)90109-9. [DOI] [PubMed] [Google Scholar]
  13. Fyfe J. A., Keller P. M., Furman P. A., Miller R. L., Elion G. B. Thymidine kinase from herpes simplex virus phosphorylates the new antiviral compound, 9-(2-hydroxyethoxymethyl)guanine. J Biol Chem. 1978 Dec 25;253(24):8721–8727. [PubMed] [Google Scholar]
  14. Garcia J. V., Bich-Thuy L. T., Stafford J., Queen C. Synergism between immunoglobulin enhancers and promoters. Nature. 1986 Jul 24;322(6077):383–385. doi: 10.1038/322383a0. [DOI] [PubMed] [Google Scholar]
  15. Gerlinger P., LeMeur M., Irrmann C., Renard P., Wasylyk C., Wasylyk B. B-lymphocyte targeting of gene expression in transgenic mice with the immunoglobulin heavy-chain enhancer. Nucleic Acids Res. 1986 Aug 26;14(16):6565–6577. doi: 10.1093/nar/14.16.6565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gillies S. D., Morrison S. L., Oi V. T., Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. doi: 10.1016/0092-8674(83)90014-4. [DOI] [PubMed] [Google Scholar]
  17. Hammer R. E., Brinster R. L., Rosenfeld M. G., Evans R. M., Mayo K. E. Expression of human growth hormone-releasing factor in transgenic mice results in increased somatic growth. 1985 May 30-Jun 5Nature. 315(6018):413–416. doi: 10.1038/315413a0. [DOI] [PubMed] [Google Scholar]
  18. Havran W. L., Poenie M., Kimura J., Tsien R., Weiss A., Allison J. P. Expression and function of the CD3-antigen receptor on murine CD4+8+ thymocytes. Nature. 1987 Nov 12;330(6144):170–173. doi: 10.1038/330170a0. [DOI] [PubMed] [Google Scholar]
  19. Landel C. P., Zhao J., Bok D., Evans G. A. Lens-specific expression of recombinant ricin induces developmental defects in the eyes of transgenic mice. Genes Dev. 1988 Sep;2(9):1168–1178. doi: 10.1101/gad.2.9.1168. [DOI] [PubMed] [Google Scholar]
  20. Lesley J., Schulte R., Hyman R. Kinetics of thymus repopulation by intrathymic progenitors after intravenous injection: evidence for successive repopulation by an IL-2R+, Pgp-1- and by an IL-2R-, Pgp-1+ progenitor. Cell Immunol. 1988 Dec;117(2):378–388. doi: 10.1016/0008-8749(88)90127-x. [DOI] [PubMed] [Google Scholar]
  21. McKnight S. L. The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene. Nucleic Acids Res. 1980 Dec 20;8(24):5949–5964. doi: 10.1093/nar/8.24.5949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miescher G. C., Howe R. C., Lees R. K., MacDonald H. R. CD3-associated alpha/beta and gamma/delta heterodimeric receptors are expressed by distinct populations of CD4- CD8- thymocytes. J Immunol. 1988 Mar 15;140(6):1779–1782. [PubMed] [Google Scholar]
  23. Palmiter R. D., Behringer R. R., Quaife C. J., Maxwell F., Maxwell I. H., Brinster R. L. Cell lineage ablation in transgenic mice by cell-specific expression of a toxin gene. Cell. 1987 Jul 31;50(3):435–443. doi: 10.1016/0092-8674(87)90497-1. [DOI] [PubMed] [Google Scholar]
  24. Palmiter R. D., Brinster R. L., Hammer R. E., Trumbauer M. E., Rosenfeld M. G., Birnberg N. C., Evans R. M. Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes. Nature. 1982 Dec 16;300(5893):611–615. doi: 10.1038/300611a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Penit C., Vasseur F. Sequential events in thymocyte differentiation and thymus regeneration revealed by a combination of bromodeoxyuridine DNA labeling and antimitotic drug treatment. J Immunol. 1988 May 15;140(10):3315–3323. [PubMed] [Google Scholar]
  26. Queen C., Baltimore D. Immunoglobulin gene transcription is activated by downstream sequence elements. Cell. 1983 Jul;33(3):741–748. doi: 10.1016/0092-8674(83)90016-8. [DOI] [PubMed] [Google Scholar]
  27. Shimonkevitz R. P., Husmann L. A., Bevan M. J., Crispe I. N. Transient expression of IL-2 receptor precedes the differentiation of immature thymocytes. Nature. 1987 Sep 10;329(6135):157–159. doi: 10.1038/329157a0. [DOI] [PubMed] [Google Scholar]
  28. St Clair M. H., Lambe C. U., Furman P. A. Inhibition by ganciclovir of cell growth and DNA synthesis of cells biochemically transformed with herpesvirus genetic information. Antimicrob Agents Chemother. 1987 Jun;31(6):844–849. doi: 10.1128/aac.31.6.844. [DOI] [PMC free article] [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