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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
. 1991 Dec 1;88(23):10505–10509. doi: 10.1073/pnas.88.23.10505

Expression of Rous sarcoma virus-derived retroviral vectors in the avian blastoderm: potential as stable genetic markers.

S T Reddy 1, A W Stoker 1, M J Bissell 1
PMCID: PMC52957  PMID: 1660139

Abstract

Retroviruses are valuable tools in studies of embryonic development, both as gene expression vectors and as cell lineage markers. In this study early chicken blastoderm cells are shown to be permissive for infection by Rous sarcoma virus and derivative replication-defective vectors, and, in contrast to previously published data, these cells will readily express viral genes. In cultured blastoderm cells, Rous sarcoma virus stably integrates and is transcribed efficiently, producing infectious virus particles. Using replication-defective vectors encoding the bacterial lacZ gene, we further show that blastoderms can be infected in culture and in ovo. In ovo, lacZ expression is seen within 24 hr of virus inoculation, and by 96 hr stably expressing clones of cells are observed in diverse tissues throughout the embryo, including epidermis, somites, and heart, as well as in extraembryonic membranes. Given the rapid onset of vector expression and the broad range of permissive cell types, it should be feasible to use Rous sarcoma virus-derived retroviruses as early lineage markers and expression vectors beginning at the blastoderm stage of avian embryogenesis.

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Selected References

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  1. Bissell M. J., Farson D., Tung A. S. Cell shape and hexose transport in normal and virus-transformed cells in culture. J Supramol Struct. 1977;6(1):1–12. doi: 10.1002/jss.400060102. [DOI] [PubMed] [Google Scholar]
  2. Bosselman R. A., Hsu R. Y., Boggs T., Hu S., Bruszewski J., Ou S., Kozar L., Martin F., Green C., Jacobsen F. Germline transmission of exogenous genes in the chicken. Science. 1989 Jan 27;243(4890):533–535. doi: 10.1126/science.2536194. [DOI] [PubMed] [Google Scholar]
  3. Bosselman R. A., Hsu R. Y., Boggs T., Hu S., Bruszewski J., Ou S., Souza L., Kozar L., Martin F., Nicolson M. Replication-defective vectors of reticuloendotheliosis virus transduce exogenous genes into somatic stem cells of the unincubated chicken embryo. J Virol. 1989 Jun;63(6):2680–2689. doi: 10.1128/jvi.63.6.2680-2689.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dolberg D. S., Bissell M. J. Inability of Rous sarcoma virus to cause sarcomas in the avian embryo. Nature. 1984 Jun 7;309(5968):552–556. doi: 10.1038/309552a0. [DOI] [PubMed] [Google Scholar]
  5. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  6. Galileo D. S., Gray G. E., Owens G. C., Majors J., Sanes J. R. Neurons and glia arise from a common progenitor in chicken optic tectum: demonstration with two retroviruses and cell type-specific antibodies. Proc Natl Acad Sci U S A. 1990 Jan;87(1):458–462. doi: 10.1073/pnas.87.1.458. [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. Howlett A. R., Carter V. C., Martin G. S., Bissell M. J. pp60v-src tyrosine kinase is expressed and active in sarcoma-free avian embryos microinjected with Rous sarcoma virus. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7587–7591. doi: 10.1073/pnas.85.20.7587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Howlett A. R., Cullen B., Hertle M., Bissell M. J. Tissue tropism and temporal expression of Rous sarcoma virus in embryonic avian limb in ovo. Oncogene Res. 1987 Aug;1(3):255–263. [PubMed] [Google Scholar]
  10. Jähner D., Jaenisch R. Integration of Moloney leukaemia virus into the germ line of mice: correlation between site of integration and virus activation. Nature. 1980 Oct 2;287(5781):456–458. doi: 10.1038/287456a0. [DOI] [PubMed] [Google Scholar]
  11. Jähner D., Jaenisch R. Retrovirus-induced de novo methylation of flanking host sequences correlates with gene inactivity. Nature. 1985 Jun 13;315(6020):594–597. doi: 10.1038/315594a0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Lee E. Y., Parry G., Bissell M. J. Modulation of secreted proteins of mouse mammary epithelial cells by the collagenous substrata. J Cell Biol. 1984 Jan;98(1):146–155. doi: 10.1083/jcb.98.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mitrani E., Coffin J., Boedtker H., Doty P. Rous sarcoma virus is integrated but not expressed in chicken early embryonic cells. Proc Natl Acad Sci U S A. 1987 May;84(9):2781–2784. doi: 10.1073/pnas.84.9.2781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Petitte J. N., Clark M. E., Liu G., Verrinder Gibbins A. M., Etches R. J. Production of somatic and germline chimeras in the chicken by transfer of early blastodermal cells. Development. 1990 Jan;108(1):185–189. doi: 10.1242/dev.108.1.185. [DOI] [PubMed] [Google Scholar]
  16. Price J., Turner D., Cepko C. Lineage analysis in the vertebrate nervous system by retrovirus-mediated gene transfer. Proc Natl Acad Sci U S A. 1987 Jan;84(1):156–160. doi: 10.1073/pnas.84.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sanes J. R., Rubenstein J. L., Nicolas J. F. Use of a recombinant retrovirus to study post-implantation cell lineage in mouse embryos. EMBO J. 1986 Dec 1;5(12):3133–3142. doi: 10.1002/j.1460-2075.1986.tb04620.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Savatier P., Morgenstern J., Beddington R. S. Permissiveness to murine leukemia, virus expression during preimplantation and early postimplantation mouse development. Development. 1990 Jul;109(3):655–665. doi: 10.1242/dev.109.3.655. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Stoker A. W., Bissell M. J. Development of avian sarcoma and leukosis virus-based vector-packaging cell lines. J Virol. 1988 Mar;62(3):1008–1015. doi: 10.1128/jvi.62.3.1008-1015.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stoker A. W., Hatier C., Bissell M. J. The embryonic environment strongly attenuates v-src oncogenesis in mesenchymal and epithelial tissues, but not in endothelia. J Cell Biol. 1990 Jul;111(1):217–228. doi: 10.1083/jcb.111.1.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]

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