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 Dec;87(23):9202–9206. doi: 10.1073/pnas.87.23.9202

Embryonic stem cell virus, a recombinant murine retrovirus with expression in embryonic stem cells.

M Grez 1, E Akgün 1, F Hilberg 1, W Ostertag 1
PMCID: PMC55132  PMID: 2251265

Abstract

The expression of Moloney murine leukemia virus and vectors derived from it is restricted in undifferentiated mouse embryonal carcinoma and embryonal stem (ES) cells. We have developed a retroviral vector, the murine embryonic stem cell virus (MESV), that is active in embryonal carcinoma and ES cells. MESV was derived from a retroviral mutant [PCC4-cell-passaged myeloproliferative sarcoma virus (PCMV)] expressed in embryonal carcinoma cells but not in ES cells. The enhancer region of PCMV was shown to be functional in both cell types, but sequences within the 5' untranslated region of PCMV were found to restrict viral expression in ES cells. Replacement of this region by related sequences obtained from the dl-587rev retrovirus results in MESV, a modified PCMV virus that confers G418 resistance to fibroblasts and ES cells with similar efficiencies. Expression of MESV in ES cells is mediated by transcriptional regulatory elements within the 5' long terminal repeat of the viral genome.

Full text

PDF

Images in this article

9203Image
on p.9203
9205Image
on p.9205
9205Image
on p.9205

Selected References

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

  1. Barklis E., Mulligan R. C., Jaenisch R. Chromosomal position or virus mutation permits retrovirus expression in embryonal carcinoma cells. Cell. 1986 Nov 7;47(3):391–399. doi: 10.1016/0092-8674(86)90596-9. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Colicelli J., Goff S. P. Isolation of a recombinant murine leukemia virus utilizing a new primer tRNA. J Virol. 1986 Jan;57(1):37–45. doi: 10.1128/jvi.57.1.37-45.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Feuer G., Taketo M., Hanecak R. C., Fan H. Two blocks in Moloney murine leukemia virus expression in undifferentiated F9 embryonal carcinoma cells as determined by transient expression assays. J Virol. 1989 May;63(5):2317–2324. doi: 10.1128/jvi.63.5.2317-2324.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Franz T., Hilberg F., Seliger B., Stocking C., Ostertag W. Retroviral mutants efficiently expressed in embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1986 May;83(10):3292–3296. doi: 10.1073/pnas.83.10.3292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Gorman C. M., Rigby P. W., Lane D. P. Negative regulation of viral enhancers in undifferentiated embryonic stem cells. Cell. 1985 Sep;42(2):519–526. doi: 10.1016/0092-8674(85)90109-6. [DOI] [PubMed] [Google Scholar]
  9. Hilberg F., Stocking C., Ostertag W., Grez M. Functional analysis of a retroviral host-range mutant: altered long terminal repeat sequences allow expression in embryonal carcinoma cells. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5232–5236. doi: 10.1073/pnas.84.15.5232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Loh T. P., Sievert L. L., Scott R. W. Proviral sequences that restrict retroviral expression in mouse embryonal carcinoma cells. Mol Cell Biol. 1987 Oct;7(10):3775–3784. doi: 10.1128/mcb.7.10.3775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
  13. Martin G. R. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7634–7638. doi: 10.1073/pnas.78.12.7634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Ostertag W., Vehmeyer K., Fagg B., Pragnell I. B., Paetz W., Le Bousse M. C., Smadja-Joffe F., Klein B., Jasmin C., Eisen H. Myeloproliferative virus, a cloned murine sarcoma virus with spleen focus-forming properties in adult mice. J Virol. 1980 Feb;33(2):573–582. doi: 10.1128/jvi.33.2.573-582.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Seliger B., Kollek R., Stocking C., Franz T., Ostertag W. Viral transfer, transcription, and rescue of a selectable myeloproliferative sarcoma virus in embryonal cell lines: expression of the mos oncogene. Mol Cell Biol. 1986 Jan;6(1):286–293. doi: 10.1128/mcb.6.1.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith A. G., Hooper M. L. Buffalo rat liver cells produce a diffusible activity which inhibits the differentiation of murine embryonal carcinoma and embryonic stem cells. Dev Biol. 1987 May;121(1):1–9. doi: 10.1016/0012-1606(87)90132-1. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Stewart C. L., Vanek M., Wagner E. F. Expression of foreign genes from retroviral vectors in mouse teratocarcinoma chimaeras. EMBO J. 1985 Dec 30;4(13B):3701–3709. doi: 10.1002/j.1460-2075.1985.tb04138.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stocking C., Kollek R., Bergholz U., Ostertag W. Long terminal repeat sequences impart hematopoietic transformation properties to the myeloproliferative sarcoma virus. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5746–5750. doi: 10.1073/pnas.82.17.5746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Taketo M., Gilboa E., Sherman M. I. Isolation of embryonal carcinoma cell lines that express integrated recombinant genes flanked by the Moloney murine leukemia virus long terminal repeat. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2422–2426. doi: 10.1073/pnas.82.8.2422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Teich N. M., Weiss R. A., Martin G. R., Lowy D. R. Virus infection of murine teratocarcinoma stem cell lines. Cell. 1977 Dec;12(4):973–982. doi: 10.1016/0092-8674(77)90162-3. [DOI] [PubMed] [Google Scholar]
  25. Thomas K. R., Capecchi M. R. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987 Nov 6;51(3):503–512. doi: 10.1016/0092-8674(87)90646-5. [DOI] [PubMed] [Google Scholar]
  26. Wagner E. F., Keller G., Gilboa E., Rüther U., Stewart C. Gene transfer into murine stem cells and mice using retroviral vectors. Cold Spring Harb Symp Quant Biol. 1985;50:691–700. doi: 10.1101/sqb.1985.050.01.085. [DOI] [PubMed] [Google Scholar]
  27. Weiher H., Barklis E., Ostertag W., Jaenisch R. Two distinct sequence elements mediate retroviral gene expression in embryonal carcinoma cells. J Virol. 1987 Sep;61(9):2742–2746. doi: 10.1128/jvi.61.9.2742-2746.1987. [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