Research Paper | Published:

Rescuing Transgene Expression by Co-Integration

Bio/Technologyvolume 10pages14501454 (1992) | Download Citation

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Abstract

To test whether foreign gene expression can be improved in transgenic mice by manipulating the site of integration, we co-integrated the efficiently expressed sheep β-lactoglobulin gene with two poorly expressed β-lactoglobulin-derived genes encoding human proteins. In each case, we observed a significant improvement in the frequency and level of expression of the hybrid gene. “Rescuing” transgene expression by co-integration may provide a general solution for improving the efficiency of heterologous gene expression in transgenic animals.

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References

  1. 1

    Simons, J.P., McClenaghan, M. and Clark, A.J. 1987. Alteration of the quality of milk by expression of sheep β-lactoglobulin in transgenic mice. Nature 328: 530–532.

  2. 2

    Harris, S., McClenaghan, M., Simons, J.P., Ali, S. and Clark, A.J. 1991. Developmental regulation of the sheep β-lactoglobulin gene in transgenic mice. Devel. Genet. 12: 299–307.

  3. 3

    Archibald, A.L., McClenaghan, M., Hornsey, V., Simons, J.P. and Clark, A.J. 1990. High level expression of biologically active human α1-antitrypsin in transgenic mice. Proc. Natl. Acad. Sci. USA 87: 5178–5182.

  4. 4

    Wright, G., Carver, A., Cottom, D., Reeves, A., Scott, A., Simons, P., Wilmut, I., Garner, I. and Colman, A. 1991. High level expression of active human α-1-antitrypsin in the milk of transgenic sheep. Bio/Technology 9: 830–834.

  5. 5

    Whitelaw, C.B.A., Archibald, A.L., Harris, S., McClenaghan, M., Simons, J.P. and Clark, A.J. 1991. Targeting expression to the mammary gland; intronic sequences can enhance the efficiency of gene expression in transgenic mice. Transgenic Res. 1 3–13.

  6. 6

    Brinster, R.L., Allen, J.M., Behringer, R.R., Gelinas, R.E. and Palmiter, R.D. 1988. Introns increase transcriptional efficiency in transgenic mice. Proc. Natl. Acad. Sci. USA 85: 836–840.

  7. 7

    Palmiter, R.D., Sandgren, E.P., Avarbock, M.R., Allen, J.M. and Brinster, R.L. 1991. Heterologous introns can increase expression in transgenic mice. Proc. Natl. Acad. Sci. USA 88: 478–482.

  8. 8

    Allen, N.D., Cran, D.G., Barton, S.G., Hettle, S., Reik, W. and Surani, M.A. 1988. Trangenes as probes for active chromosomal domains in mouse development. Nature 333: 852–855.

  9. 9

    Al-Shawi, R., Kinnaird, J., Burke, J. and Bishop, J.O. 1990. Expression of a foreign gene in a line of transgenic mice is modulated by a chromosomal position effect. Mol. Cell. Biol. 10: 1192–1198.

  10. 10

    Bonnerot, C., Grimber, G., Briand, P. and Nicolas, J.-F. 1990. Patterns of expression of position dependent integrated transgenes in mouse embryos. Proc. Natl. Acad. Sci. USA 87: 6331–6335.

  11. 11

    Storb, U., Pinkert, C., Arp, B., Engler, P., Gollahon, K., Manz, J., Brady, W. and Brinster, R.L. 1986. Transgenic mice with mu and kappa genes encoding antiphosphorylcholine. J. Exp. Med. 164: 627–641.

  12. 12

    Behringer, R.R., Ryan, T.M., Reilly, M.P., Asakura, T., Palmiter, R.D. and Brinster, R.L., 1989. Synthesis of functional hemoglobin in transgenic mice. Science 245: 971–973.

  13. 13

    Whitelaw, C.B.A., Harris, S., McClenaghan, M., Simons, J.P. and Clark, A.J. 1992. Position-independent expression of the ovine β-lactoglobulin gene in transgenic mice. Biochem. J. 286 31–39.

  14. 14

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

  15. 15

    Kellum, R. and Schedl, P. 1991. A position effect assay for boundaries of higher order chromosomal domains. Cell 64: 941–950.

  16. 16

    Ali, S. and Clark, A.J. 1988. Characterisation of the gene encoding ovine β-lactoglobulin. J. Mol. Biol. 199: 415–426.

  17. 17

    Ali, S., McClenaghan, M., Simons, J.P. and Clark, A.J. 1990. Characterisation of the alleles encoding ovine β-lactoglobulins A and B. Gene 91: 201–207.

  18. 18

    Clark, A.J., Bessos, H., Bishop, J.O., Brown, P., Harris, S., Lathe, R., McClenaghan, M., Prowse, C., Simons, J.P., Whitelaw, C.B.A. and Wilmut, I. 1989. Expression of human anti-hemophilic factor IX in the milk of transgenic sheep. Bio/Technology 7: 487–492.

  19. 19

    Ciliberto, G., Dente, L. and Cortese, R. 1985. Cell-specific expression of a transfected α1-antirypsin gene. Cell 41: 531–540.

  20. 20

    Anson, D.S., Austen, D.E.G. and Brownlee, G.G. 1985. Expression of active human clotting factor IX from recombinant DNA clones in mammalian cells. Nature 315: 683–685.

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Author notes

  1. A. J. Clark: Corresponding author.

Affiliations

  1. AFRC Institute of Animal Physiology and Genetics Research, Edinburgh Research Station, Roslin, Midlothian, EH25 9PS, Scotland

    • A. J. Clark
    • , A. Cowper
    • , R. Wallace
    •  & J. P. Simons
  2. Pharmaceutical Proteins Ltd, Centre for Genome Research, King's Buildings, West Mains Road, Edinburgh, EH9 3JQ, Scotland

    • G. Wright

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DOI

https://doi.org/10.1038/nbt1192-1450

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