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. 1985 Dec 30;4(13B):3701–3709. doi: 10.1002/j.1460-2075.1985.tb04138.x

Expression of foreign genes from retroviral vectors in mouse teratocarcinoma chimaeras.

C L Stewart, M Vanek, E F Wagner
PMCID: PMC554721  PMID: 2419128

Abstract

The formation of chimaeric mice from embryonic stem cells (EC and ES) carrying and exposing foreign genes is described. Two retroviral vectors were used to stably introduce genes, by virus infection and selection into EC and ES cells. In the first series EC cell clones were isolated that contained a single intact copy of the vector, which expressed the neomycin resistance gene (neo) from the 5' long terminal repeat (LTR). Following the formation of chimaeras with one of the clones, expression of the foreign gene was found in all chimaeric tissues examined. In the second series, a vector was used which contained the OK10 v-myc oncogene under the control of the 5' LTR as well as the neo gene expressed from an internal thymidine kinase (TK) promoter. ES cell clones were again isolated and used to form chimaeras. Although expression of the TK neo transcripts was maintained in the chimaeric tissues analysed, no expression of the v-myc transcripts from the 5' LTR was detected. These results show that genes can be introduced into mice, using clones of embryonic stem cells selected in vitro and that the expression of the genes can be maintained probably throughout development and in all tissues. The advantages this approach offers compared with DNA injection into eggs and the use of retroviruses as vectors for introducing genes into mice are discussed.

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Selected References

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

  1. Bradley A., Evans M., Kaufman M. H., Robertson E. Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature. 1984 May 17;309(5965):255–256. doi: 10.1038/309255a0. [DOI] [PubMed] [Google Scholar]
  2. Chada K., Magram J., Raphael K., Radice G., Lacy E., Costantini F. Specific expression of a foreign beta-globin gene in erythroid cells of transgenic mice. 1985 Mar 28-Apr 3Nature. 314(6009):377–380. doi: 10.1038/314377a0. [DOI] [PubMed] [Google Scholar]
  3. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. Costantini F., Lacy E. Introduction of a rabbit beta-globin gene into the mouse germ line. Nature. 1981 Nov 5;294(5836):92–94. doi: 10.1038/294092a0. [DOI] [PubMed] [Google Scholar]
  5. Dewey M. J., Martin D. W., Jr, Martin G. R., Mintz B. Mosaic mice with teratocarcinoma-derived mutant cells deficient in hypoxanthine phosphoribosyltransferase. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5564–5568. doi: 10.1073/pnas.74.12.5564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duboule D., Croce C. M., Illmensee K. Tissue preference and differentiation of malignant rat x mouse hybrid cells in chimaeric mouse fetuses. EMBO J. 1982;1(12):1595–1603. doi: 10.1002/j.1460-2075.1982.tb01361.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eicher E. M., Washburn L. L. Assignment of genes to regions of mouse chromosomes. Proc Natl Acad Sci U S A. 1978 Feb;75(2):946–950. doi: 10.1073/pnas.75.2.946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Evans M. J., Kaufman M. H. Establishment in culture of pluripotential cells from mouse embryos. Nature. 1981 Jul 9;292(5819):154–156. doi: 10.1038/292154a0. [DOI] [PubMed] [Google Scholar]
  9. Gautsch J. W., Wilson M. C. Delayed de novo methylation in teratocarcinoma suggests additional tissue-specific mechanisms for controlling gene expression. Nature. 1983 Jan 6;301(5895):32–37. doi: 10.1038/301032a0. [DOI] [PubMed] [Google Scholar]
  10. Gordon J. W., Ruddle F. H. Gene transfer into mouse embryos: production of transgenic mice by pronuclear injection. Methods Enzymol. 1983;101:411–433. doi: 10.1016/0076-6879(83)01031-9. [DOI] [PubMed] [Google Scholar]
  11. Gordon J. W., Scangos G. A., Plotkin D. J., Barbosa J. A., Ruddle F. H. Genetic transformation of mouse embryos by microinjection of purified DNA. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7380–7384. doi: 10.1073/pnas.77.12.7380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grosschedl R., Weaver D., Baltimore D., Costantini F. Introduction of a mu immunoglobulin gene into the mouse germ line: specific expression in lymphoid cells and synthesis of functional antibody. Cell. 1984 Oct;38(3):647–658. doi: 10.1016/0092-8674(84)90259-9. [DOI] [PubMed] [Google Scholar]
  13. Groudine M., Eisenman R., Weintraub H. Chromatin structure of endogenous retroviral genes and activation by an inhibitor of DNA methylation. Nature. 1981 Jul 23;292(5821):311–317. doi: 10.1038/292311a0. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Hanahan D. Heritable formation of pancreatic beta-cell tumours in transgenic mice expressing recombinant insulin/simian virus 40 oncogenes. Nature. 1985 May 9;315(6015):115–122. doi: 10.1038/315115a0. [DOI] [PubMed] [Google Scholar]
  16. Harbers K., Schnieke A., Stuhlmann H., Jähner D., Jaenisch R. DNA methylation and gene expression: endogenous retroviral genome becomes infectious after molecular cloning. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7609–7613. doi: 10.1073/pnas.78.12.7609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jaenisch R., Fan H., Croker B. Infection of preimplantation mouse embryos and of newborn mice with leukemia virus: tissue distribution of viral DNA and RNA and leukemogenesis in the adult animal. Proc Natl Acad Sci U S A. 1975 Oct;72(10):4008–4012. doi: 10.1073/pnas.72.10.4008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jaenisch R., Jähner D., Nobis P., Simon I., Löhler J., Harbers K., Grotkopp D. Chromosomal position and activation of retroviral genomes inserted into the germ line of mice. Cell. 1981 May;24(2):519–529. doi: 10.1016/0092-8674(81)90343-3. [DOI] [PubMed] [Google Scholar]
  19. Jaenisch R., Mintz B. Simian virus 40 DNA sequences in DNA of healthy adult mice derived from preimplantation blastocysts injected with viral DNA. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1250–1254. doi: 10.1073/pnas.71.4.1250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jähner D., Stuhlmann H., Stewart C. L., Harbers K., Löhler J., Simon I., Jaenisch R. De novo methylation and expression of retroviral genomes during mouse embryogenesis. Nature. 1982 Aug 12;298(5875):623–628. doi: 10.1038/298623a0. [DOI] [PubMed] [Google Scholar]
  21. Keller G., Paige C., Gilboa E., Wagner E. F. Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors. Nature. 1985 Nov 14;318(6042):149–154. doi: 10.1038/318149a0. [DOI] [PubMed] [Google Scholar]
  22. Kondoh H., Takahashi Y., Okada T. S. Differentiation-dependent expression of the chicken delta-crystallin gene introduced into mouse teratocarcinoma stem cells. EMBO J. 1984 Sep;3(9):2009–2014. doi: 10.1002/j.1460-2075.1984.tb02083.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lacy E., Roberts S., Evans E. P., Burtenshaw M. D., Costantini F. D. A foreign beta-globin gene in transgenic mice: integration at abnormal chromosomal positions and expression in inappropriate tissues. Cell. 1983 Sep;34(2):343–358. doi: 10.1016/0092-8674(83)90369-0. [DOI] [PubMed] [Google Scholar]
  24. Levis R., Hazelrigg T., Rubin G. M. Effects of genomic position on the expression of transduced copies of the white gene of Drosophila. Science. 1985 Aug 9;229(4713):558–561. doi: 10.1126/science.2992080. [DOI] [PubMed] [Google Scholar]
  25. Linney E., Davis B., Overhauser J., Chao E., Fan H. Non-function of a Moloney murine leukaemia virus regulatory sequence in F9 embryonal carcinoma cells. 1984 Mar 29-Apr 4Nature. 308(5958):470–472. doi: 10.1038/308470a0. [DOI] [PubMed] [Google Scholar]
  26. Löhler J., Timpl R., Jaenisch R. Embryonic lethal mutation in mouse collagen I gene causes rupture of blood vessels and is associated with erythropoietic and mesenchymal cell death. Cell. 1984 Sep;38(2):597–607. doi: 10.1016/0092-8674(84)90514-2. [DOI] [PubMed] [Google Scholar]
  27. McBurney M. W., Strutt B. J. Genetic activity of X chromosomes in pluripotent female teratocarcinoma cells and their differentiated progeny. Cell. 1980 Sep;21(2):357–364. doi: 10.1016/0092-8674(80)90472-9. [DOI] [PubMed] [Google Scholar]
  28. Niwa O., Yokota Y., Ishida H., Sugahara T. Independent mechanisms involved in suppression of the Moloney leukemia virus genome during differentiation of murine teratocarcinoma cells. Cell. 1983 Apr;32(4):1105–1113. doi: 10.1016/0092-8674(83)90294-5. [DOI] [PubMed] [Google Scholar]
  29. Ornitz D. M., Palmiter R. D., Hammer R. E., Brinster R. L., Swift G. H., MacDonald R. J. Specific expression of an elastase-human growth hormone fusion gene in pancreatic acinar cells of transgenic mice. Nature. 1985 Feb 14;313(6003):600–602. doi: 10.1038/313600a0. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Papaioannou V. E., McBurney M. W., Gardner R. L., Evans M. J. Fate of teratocarcinoma cells injected into early mouse embryos. Nature. 1975 Nov 6;258(5530):70–73. doi: 10.1038/258070a0. [DOI] [PubMed] [Google Scholar]
  32. Pellicer A., Wagner E. F., el-Kareh A., Dewey M. J., Reuser A. J., Silverstein S., Axel R., Mintz B. Introduction of a viral thymidine kinase gene and the human beta-globin gene into developmentally multipotential mouse teratocarcinoma cells. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2098–2102. doi: 10.1073/pnas.77.4.2098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reiss B., Sprengel R., Will H., Schaller H. A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene. 1984 Oct;30(1-3):211–217. doi: 10.1016/0378-1119(84)90122-7. [DOI] [PubMed] [Google Scholar]
  34. Rosenthal A., Wright S., Cedar H., Flavell R., Grosveld F. Regulated expression of an introduced MHC H-2K bm1 gene in murine embryonal carcinoma cells. Nature. 1984 Aug 2;310(5976):415–418. doi: 10.1038/310415a0. [DOI] [PubMed] [Google Scholar]
  35. Rubenstein J. L., Nicolas J. F., Jacob F. Construction of a retrovirus capable of transducing and expressing genes in multipotential embryonic cells. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7137–7140. doi: 10.1073/pnas.81.22.7137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schnieke A., Harbers K., Jaenisch R. Embryonic lethal mutation in mice induced by retrovirus insertion into the alpha 1(I) collagen gene. 1983 Jul 28-Aug 3Nature. 304(5924):315–320. doi: 10.1038/304315a0. [DOI] [PubMed] [Google Scholar]
  37. Scott R. W., Vogt T. F., Croke M. E., Tilghman S. M. Tissue-specific activation of a cloned alpha-fetoprotein gene during differentiation of a transfected embryonal carcinoma cell line. Nature. 1984 Aug 16;310(5978):562–567. doi: 10.1038/310562a0. [DOI] [PubMed] [Google Scholar]
  38. Sorge J., Cutting A. E., Erdman V. D., Gautsch J. W. Integration-specific retrovirus expression in embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6627–6631. doi: 10.1073/pnas.81.21.6627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  40. Stewart C. L. Formation of viable chimaeras by aggregation between teratocarcinomas and preimplantation mouse embryos. J Embryol Exp Morphol. 1982 Feb;67:167–179. [PubMed] [Google Scholar]
  41. Stewart C. L., Stuhlmann H., Jähner D., Jaenisch R. De novo methylation, expression, and infectivity of retroviral genomes introduced into embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4098–4102. doi: 10.1073/pnas.79.13.4098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stewart C. Aggregation between teratocarcinoma cells and preimplantation mouse embryos. J Embryol Exp Morphol. 1980 Aug;58:289–302. [PubMed] [Google Scholar]
  43. Stewart T. A., Mintz B. Successive generations of mice produced from an established culture line of euploid teratocarcinoma cells. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6314–6318. doi: 10.1073/pnas.78.10.6314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Stewart T. A., Pattengale P. K., Leder P. Spontaneous mammary adenocarcinomas in transgenic mice that carry and express MTV/myc fusion genes. Cell. 1984 Oct;38(3):627–637. doi: 10.1016/0092-8674(84)90257-5. [DOI] [PubMed] [Google Scholar]
  45. Stuhlmann H., Cone R., Mulligan R. C., Jaenisch R. Introduction of a selectable gene into different animal tissue by a retrovirus recombinant vector. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7151–7155. doi: 10.1073/pnas.81.22.7151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Vennström B., Kahn P., Adkins B., Enrietto P., Hayman M. J., Graf T., Luciw P. Transformation of mammalian fibroblasts and macrophages in vitro by a murine retrovirus encoding an avian v-myc oncogene. EMBO J. 1984 Dec 20;3(13):3223–3229. doi: 10.1002/j.1460-2075.1984.tb02282.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wagner E. F., Mintz B. Transfer of nonselectable genes into mouse teratocarcinoma cells and transcription of the transferred human beta-globin gene. Mol Cell Biol. 1982 Feb;2(2):190–198. doi: 10.1128/mcb.2.2.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wagner E. F., Stewart T. A., Mintz B. The human beta-globin gene and a functional viral thymidine kinase gene in developing mice. Proc Natl Acad Sci U S A. 1981 Aug;78(8):5016–5020. doi: 10.1073/pnas.78.8.5016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wagner E. F., Vanek M., Vennström B. Transfer of genes into embryonal carcinoma cells by retrovirus infection: efficient expression from an internal promoter. EMBO J. 1985 Mar;4(3):663–666. doi: 10.1002/j.1460-2075.1985.tb03680.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. van Someren H., Beijersbergen van Henegouwen H., Los W., Wurzer-Figurelli E., Doppert B., Vervloet M., Meera Khan P. Enzyme electrophoresis on cellulose acetate gel. II. Zymogram patterns in man-Chinese hamster somatic cell hybrids. Humangenetik. 1974;25(3):189–201. doi: 10.1007/BF00281426. [DOI] [PubMed] [Google Scholar]

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