, Volume 31, Issue 1–2, pp 3–8 | Cite as

The changing role of cell culture in the generation of transgenic livestock

  • C. B. A. Whitelaw
  • E. Farini
  • J. Webster


Transgenesis may allow the generation of farm animals with altered phenotype, animal models for research and animal bioreactors. Although such animals have been produced, the time and expense involved in generating transgenic livestock and then evaluating the transgene expression pattern is very restrictive. If questions about the ability and efficiency of expression could be asked solely in vitro rapid progress could be achieved. Unfortunately, experiments addressing transcriptional control in vitro have proved unreliable in their ability to indicate whether a transgene will be transcribed or not. However, initial studies suggest that cell culture may be able to predict in vivo post-transcriptional events. We review these issues and propose that strategies which engineer the transgene integration site could enhance the probability for efficient expression. This approach has now become feasible with the development of techniques allowing animals to be generated from somatic cells by nuclear transfer. The important step in this procedure is the use of cells grown in culture as the source of genetic information, allowing the selection of specific transgene integration events. This technology which has dramatically increased the potential use of transgenic livestock for both agricultural and biotechnological applications, is based on standard cell culture methodology. We are now at the start of a new era in large animal transgenics.

cell culture livestock milk nuclear transfer transgenic 


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  1. Al-Shawi R, Kinnaird J, Burke J and Bishop JO (1990) Expression of a foreign gene in a line of mice is modulated by chromosomal position effect. Mol Cell Biol 10: 1192–1198.Google Scholar
  2. Ashe HL, Monks J, Wijgerde M, Fraser P and Proudfoot NJ (1997) Intergenic transcription and transduction of the human β-globin locus. Genes Dev 11: 2494–2509.Google Scholar
  3. Brinster RL, Allen JM, Behringer RR, Gelinas RE and Palmiter RD (1988) Introns increase transcriptional efficiency in transgenic mice. Proc Natl Acad Sci USA 85: 863–840.CrossRefGoogle Scholar
  4. Bronson SK, Plaetn EG, Kluckman JR, Hagaman JR, Maeda N and Smithies O (1996) Single copy transgenic mice with chosen site integration. Proc Natl Acad Sci USA 93: 9067–9072.CrossRefGoogle Scholar
  5. Burdon TG, Demmer J, Clark AJ and Watson, CJ (1994) The mammary factor MPBF is a prolactin-induced transcriptional regulator which binds to STAT factor recognition sites. FEBS Lett 350: 177–182.CrossRefGoogle Scholar
  6. Campbell KHS, McWhir J, Ritchie WA and Wilmut I (1996) Sheep cloned by nuclear transfer from a cultured cell line. Nature 380: 64–66.CrossRefGoogle Scholar
  7. Clark AJ, Bissinger P, Bullock DW, Dmak S, Wallace R, Whitelaw CBA and Yull F (1994) Chromosomal position effects and the modulation of transgene expression. Reprod Fert Dev 6: 589–598.CrossRefGoogle Scholar
  8. Dobie KW, Lee M, Fantes JA, Graham E, Clark AJ, Springbett A, Lathe R and McClenaghan M (1996) Variegated transgene expression in mouse mammary gland is determined by the transgene integration locus. Proc Natl Acad Sci USA 93: 6659–6664.CrossRefGoogle Scholar
  9. Donofrio G, Bignetti E, Clark AJ and Whitelaw CBA (1996) Comparable processing of β-lactoglobulin pre-mRNA in cell culture and transgenic mouse models. Mol Gen Genet 252: 465–469.Google Scholar
  10. Dorer DR (1997) Do transgene arrays form heterochromatin in vertebrates? Trans Res 6: 3–10.CrossRefGoogle Scholar
  11. Drews R, Paleyanda RK, Lee TK, Chang RR, Rehemtulla A, Kaufman RJ, Drohan WN and Lubon H (1995) Proteolytic maturation of protein C upon engineering the mouse mammary gland to express furin. Proc Natl Acad Sci USA 92: 10462–10466.CrossRefGoogle Scholar
  12. Farini E and Whitelaw CBA (1995) Ectopic expression of β-lactoglobulin transgenes. Mol Gen Genet 246: 734–738.CrossRefGoogle Scholar
  13. Garrick D, Fiering S, Martin DIK and Whitelaw E (1997) Repeat-induced gene silencing in mammals. Nat Genet 18: 56–59.CrossRefGoogle Scholar
  14. Guy L-G, Kothary R and Wall L (1997) Position effects in mice carrying a lacZ transgene in cis with the β-globin LCR can be explained by a graded model. Nucl Acids Res 25: 4400–4407.CrossRefGoogle Scholar
  15. Hammer RE, Pursel VG, Rexroad C, Wall RJ, Bolt DJ, Ebert KM, Palmiter RD and Brinster RL (1985) Production of transgenic rabbits, sheep and pigs by microinjection. Nature 315: 680–683.CrossRefGoogle Scholar
  16. Haynes JI, Gopal-Srivastava R and Piatigorsky J (1997) αB-crystallin TATA sequence mutations: lens-preference for the proximal TATA box and the distal TATA-like sequence in transgenic mice. Biochem Biophys Res Comm 241: 407–413.CrossRefGoogle Scholar
  17. Kornberg RD (1996) RNA polymerase II transcription control. Trends Biochem Sci 21: 325–326.CrossRefGoogle Scholar
  18. Lesueur L, Edery M, Paly J, Clark J, Kelly PA and Djiane J (1990) Prolactin stimulates milk protein promoter in CHO cells cotransfected with prolactin receptor cDNA. Mol Cell Endocrinol 71: R7-R12.CrossRefGoogle Scholar
  19. McWhir J, Schnieke A, Ansell R, Wallace H, Colman A, Scott AR and Kind AJ (1996) Selective ablation of differentiated cells permits isolation of ES cells from murine embryos with a non-permissive genetic background. Nat Genet 14: 223–226.CrossRefGoogle Scholar
  20. Melton DW, Ketchen AM and Selfridge J (1997) Stability of HPRT marker gene expression at different gene-targeted loci: observing and overcoming a position effect. Nucl Acids Res 25: 23937–3943.CrossRefGoogle Scholar
  21. Millonig JH, Emerson JA, Levorse JM and Tilghman SM (1995) Molecular analysis of the distal enhancer of the mouse α-fetoprotein gene. Mol Cell Biol 15: 3848–3856.Google Scholar
  22. Mulder LCF, Rossini M and Mora M (1997) Human CD46 aberrant splicing in transgenic mice. Gene 186: 83–86.CrossRefGoogle Scholar
  23. Petitclerc D, Attal J, Theron MC, Bearzotti M, Bolifraud P, Kann G, Stinnakre M-G, Pointu H, Puissant C and Houdebine L-M (1995) The effect of various introns and transcription terminators on the efficiency of expression vectors in various cultured cell lines and in the mammary gland of transgenic mice. J Biotechnol 40: 169–178.CrossRefGoogle Scholar
  24. Robertson G, Garrick D, Wu W, Kearns M, Martin D and Whitelaw E (1995) Position-dependent variegation of globin transgene expression in mice. Proc Natl Acad Sci USA 92: 5371–5375.CrossRefGoogle Scholar
  25. Saavedra C, Felber B and Izaurralde E (1997) The simian retrovirus-1 constitutive transport element, unlike the HIV-1 RRE, uses factors required for cellular mRNA export. Curr Biol 7: 619–628.CrossRefGoogle Scholar
  26. Seipelt RL, Spear BT, Spear E, Snow, EC and Peterson ML (1998) A non-immunoglobulin transgene and the endogenous immunoglobulin μ gene are coordinately regulated by alternative RNA processing during B-cell maturation. Mol Cell Biol 18: 1042–1048.Google Scholar
  27. Schnieke AE, Kind AJ, Ritchie WA, Mycock K, Scott AR, Ritchie M, Wilmut I, Colman A and Campbell KHS (1997) Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts. Science 278: 2130–2133.CrossRefGoogle Scholar
  28. Stacey A, Schnieke A, Kerr M, Scott A, McKee C, Cottingham I, Binas B, Wilde C and Colman A (1995) Lactation is disrupted by α-lactalbumin deficiency and can be restored by human α-lactalbumin gene replacement. Proc Natl Acad Sci USA 92: 2835–2839.CrossRefGoogle Scholar
  29. Thompson S, Clarke AR, Pow AM, Hooper ML and Melton DW (1989) Germline transmission and expression of a corrected HPRT gene produced by gene targeting in ES cells. Cell 56: 313–321.CrossRefGoogle Scholar
  30. Walters MC, Fiering S, Eidemiller J, Magis W, Groudine M and Martin DIK (1995) Enhancers increase the probability but not the level of gene expression. Proc Natl Acad Sci USA 92: 7125–7129.CrossRefGoogle Scholar
  31. Webster J, Donofrio G, Wallace R, Clark AJ and Whitelaw CBA (1997) Intronic sequences modulate the sensitivity of BLG transgenes to position effects. Gene 193: 239–343.CrossRefGoogle Scholar
  32. Webster J, Wallace RM, Clark AJ and Whitelaw CBA (1995) Tissue-specific, temporally regulated expression mediated by the proximal ovine BLG promoter in transgenic mice. Mol Cell Biol Res 41: 11–154.Google Scholar
  33. Whitelaw CBA, Harris S, McClenaghan M, Simons JP and Clark AJ (1992) Position-independent expression of the ovine BLG gene transgenic mice. Biochem J 286: 31–39.Google Scholar
  34. Wilmut I, Schnieke AE, McWhir J, Kind AJ and Campbell KHS (1997) Viable offspring derived from fetal and adult mammalian cells. Nature 385: 810–813.CrossRefGoogle Scholar
  35. Wilmut I and Whitelaw CBA (1994) Strategies for production of pharmaceutical proteins in milk. Reprod Fert Dev 6: 625–630.CrossRefGoogle Scholar
  36. Yull F, Harold G, Wallace R, Cowper A, Percy J, Cottingham I and Clark AJ (1995) Fixing human factor IX (fIX): correction of a cryptic RNA splice enables the production of biologically active fIX in the mammary gland of transgenic mice. Proc Natl Acad Sci USA 92: 10899–10903.CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • C. B. A. Whitelaw
  • E. Farini
  • J. Webster

There are no affiliations available

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