Research | Published:

Transgenic Livestock as Bioreactors: Stable Expression of Human Alpha-1-Antitrypsin by a Flock of Sheep

Nature Biotechnology volume 11, pages 12631270 (1993) | Download Citation

Subjects

Abstract

We have previously described the generation of transgenic sheep expressing human alpha 1-antitrypsin (hα1AT) in their milk. Here, we report the fidelity of transgene transmission and expression by these animals and their progeny. Transgene transmission has been demonstrated in four of six ovine lines studied. Three of these four lines have exhibited stable transmission of the transgene, whereas the fourth has produced some offspring with reduced copy numbers. Sequential lactations of founder animals has yielded very similar levels of hα1AT protein in milk. Moreover, in one line, derived from a founder male, a flock of seven Gl ewes have yielded comparable levels of hα1 AT protein in first and second lactation milk. Two G2 ewes of this line have also produced levels of human protein equivalent to their mother. Although the inheritance of the same transgene in mice was reminiscent of the situation in sheep, stable expression was observed in only one of four lines studied. The importance of these observations to the use of transgenic livestock as bioreactors for the production of human proteins is discussed.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , and 1986. Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor Laboratory Press, NY.

  2. 2.

    and 1981. Establishment in culture of pluripotential cells from mouse embryos. Nature, 292: 154–156.

  3. 3.

    and 1987. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. cell 51: 503–512.

  4. 4.

    1989. The new mouse genetics: Altering the genome by gene targeting. Trends in Genetics 5: 70–76.

  5. 5.

    , , and 1992. Modifying the mouse: design and desire. Bio/Technology 10: 534–539.

  6. 6.

    , , and 1990. The chicken lysozyme matrix attachment region increases transcription from a heterologous promoter in heterologous cells and dampens position effects on the expression of transfected genes. Mol. cell Biol. 10: 2302–2307.

  7. 7.

    , , and 1987. Position independent high level expression of the human β-Globin gene in transgenic mice. cell 51: 975–985.

  8. 8.

    , , , , , , , , and 1992. Production of functional human hemoglobin in transgenic swine. Bio/Technology 10: 557–559.

  9. 9.

    , , , , , , , , , and 1989. Expression of human anti-hemophilic factor IX in the milk of transgenic sheep. Bio/Technology 7: 487–492.

  10. 10.

    , , , , , , , and 1991. High level expression of active human α1 antitrypsin in the milk of transgenic sheep. Bio/Technology 9: 830–834.

  11. 11.

    , , , , , , , , and 1991. Transgenic production of a variant of human tissue-type plasminogen activator in goat milk: generation of transgenic goats and analysis of expression. Bio/Technology 9: 835–838.

  12. 12.

    , , , , and 1988. Gene transfer into sheep. Bio/Technology 6: 179–183.

  13. 13.

    , , , , , , , , , , , , , , and 1992. Expression of human α1 antitrypsin in transgenic sheep. Cytotechnology 9: 77–84.

  14. 14.

    , and 1990. Imprinting by DNA methylation: from transgenes to endogenous gene sequences. Development Supplement: 99–106.

  15. 15.

    and 1981. Integration and stable germline transmission of genes injected into mouse pronuclei. Science 214: 1244–1246.

  16. 16.

    , , and 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.

  17. 17.

    , , , and 1992. Matrix-attachment regions can impart position-independent regulation of a tissue-specific gene in transgenic mice. Proc. Natl. Acad. Sci. USA 89: 6943–6947.

  18. 18.

    , , , , , , , and 1992. High-level expression of heterologous protein in the milk of transgenic swine using the cDNA encoding human Protein C. Proc. Natl. Acad. Sci. USA 89: 12003–12007.

  19. 19.

    , , , and 1988. Introns increase transcriptional efficiency in transgenic mice. Proc. Natl. Acad. Sci. USA 85: 836–840.

  20. 20.

    , , , and 1991. Heterologous introns can increase expression in transgenic mice. Proc. Natl. Acad. Sci. USA 88: 478–482.

  21. 21.

    , , , and 1992. Rescuing transgene expression by co-integration. Bio/Technology 10: 1450–1454.

  22. 22.

    , , , , , , and 1991. Expression of biologically active heterodimeric bovine follicle-stimulating hormone in milk of transgenic mice. Proc. Natl. Acad. Sci. USA 88: 8327–8331.

Download references

Author information

Author notes

    • Ian Garner

    Corresponding author.

Affiliations

  1. Pharmaceutical Proteins Limited, Roslin, Midlothian, EH25 9PP, United Kingdom.

    • Andrew S. Carver
    • , Michael A. Dalrymple
    • , Gordon Wright
    • , Denise S. Cottom
    • , Dawn B. Reeves
    • , Yvonne H. Gibson
    • , Jayne L. Keenan
    • , J. David Barrass
    • , Ann R. Scott
    • , Alan Colman
    •  & Ian Garner

Authors

  1. Search for Andrew S. Carver in:

  2. Search for Michael A. Dalrymple in:

  3. Search for Gordon Wright in:

  4. Search for Denise S. Cottom in:

  5. Search for Dawn B. Reeves in:

  6. Search for Yvonne H. Gibson in:

  7. Search for Jayne L. Keenan in:

  8. Search for J. David Barrass in:

  9. Search for Ann R. Scott in:

  10. Search for Alan Colman in:

  11. Search for Ian Garner in:

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nbt1193-1263

Further reading