Advertisement

YAC Protocols pp 281-292 | Cite as

YAC Transfer into Mammalian Cells by Cell Fusion

  • Nicholas P. Davies
  • Clare Huxley
Part of the Methods in Molecular Biology™ book series (MIMB, volume 54)

Abstract

The large regions of DNA that can be cloned in yeast artificial chromosomes (YACs) are ideal for expression studies of the complex genes and gene clusters found in the mammalian genome. Such studies require that the YAC of interest be transferred into a suitable expression system, such as mammalian cells in tissue culture or transgenic animals. Recent experiments indicate that large genes cloned on YACs may be transferred intact and are often expressed at a level comparable to the endogenous genes and in a fully controlled fashion owing to the large amount of flanking DNA containing long range controlling elements (reviewed in ref. 1).

Keywords

Mammalian Cell Selectable Marker Yeast Artificial Chromosome Extrachromosomal Element Rodent Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Huxley, C. (1994) Transfer of YACs to mammalian cells and transgenic mice, in Genetic Engineering (Setlow, J. K., ed.), Plenum, New York, pp. 65–91.Google Scholar
  2. 2.
    Davies, N. P., Rosewell, I. R., Richardson, J. C., Cook, G. P., Neuberger, M. S., Brownstein, B. H., et al. (1993) Creation of mice expressing human antibody light chains by introduction of a yeast artificial chromosome containing the core region of the human immunoglobulin κ locus. BioTechnology 11, 911–914.PubMedCrossRefGoogle Scholar
  3. 3.
    Jakobovits, A., Moore, A. L., Green, L. L., Vergara, G. J., Maynard-Currie, C. E., Austin, H. A., and Klapholz, S. (1993) Germ-line transmission and expression of a human-derived yeast artificial chromosome. Nature 362, 255–258.PubMedCrossRefGoogle Scholar
  4. 4.
    Green, L. L., Hardy, M. C., Maynard-Currie, C. E., Tsuda, H., Louie, D. M., Mendez, M. J., et al. (1994) Antigen-specific human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs. Nature Genet. 7, 13–21.PubMedCrossRefGoogle Scholar
  5. 5.
    Allshire, R. C., Cranston, G., Gosden, J. R., Maule, J. C., Hastie, N. D., and Fantes, P. A. (1987) A fission yeast chromosome can replicate autonomously in mouse cells. Cell 50, 391–403.PubMedCrossRefGoogle Scholar
  6. 6.
    Ward, M., Scott, R. J., Davey, M. R., Clothier, R. H., Cocking, E. C., and Balls, M. (1986) Transfer of antibiotic resistance genes between yeast and mammalian cells under conditions favoring cell fusion. Somat. Cell. Mol. Genet. 12, 101–109.PubMedCrossRefGoogle Scholar
  7. 7.
    Traver, C. N., Klapholz, S., Hyman, R. W., and Davis, R. W. (1989) Rapid screening of a human genomic library in yeast artificial chromosomes for single-copy sequences. Proc. Natl. Acad. Sci. USA 86, 5898–5902.PubMedCrossRefGoogle Scholar
  8. 8.
    Pachnis, V., Pevny, L., Rothstein, R., and Costantini, F. (1990) Transfer of a yeast artificial chromosome carrying human DNA from Saccharomyces cerevisiae into mammalian cells. Proc. Natl. Acad. Sci. USA 87, 5109–5113.PubMedCrossRefGoogle Scholar
  9. 9.
    Pavan, W. J., Hieter, P., and Reeves, R. H. (1990) Modification and transfer into an embryonal carcinoma cell line of a 360-kilobase human-derived yeast artificial chromosome. Mol. Cell. Biol. 10, 4163–4169.PubMedGoogle Scholar
  10. 10.
    Gnirke, A., Barnes, T. S., Patterson, D., Schild, D., Featherstone, T., and Olson, M. V. (1991) Cloning and in vivo expression of the human GART gene using yeast artificial chromosomes. EMBO J. 10, 1629–1634.PubMedGoogle Scholar
  11. 11.
    Huxley, C., Hagino, Y., Schlessinger, D., and Olson, M. V. (1991) The human HPRT gene on a yeast artificial chromosome is functional when transferred to mouse cells by cell fusion. Genomics 9, 742–750.PubMedCrossRefGoogle Scholar
  12. 12.
    Davies, N. P., Rosewell, I. R., and Bruggemann, M. (1992) Targeted alterations in yeast artificial chromosomes for inter-species gene transfer. Nucleic Acids Res. 20, 2693–2698.PubMedCrossRefGoogle Scholar
  13. 13.
    Riley, J. H., Morten, J. E. N., and Anand, R. (1992) Targeted integration of neomycin into yeast artificial chromosomes (YACs) for transfection into mammalian cells. Nucleic Acids Res. 20, 2971–2976.PubMedCrossRefGoogle Scholar
  14. 14.
    Demmer, L. A. and Chaplin, D. D. (1993) Simultaneous transfer of four functional genes from the HLA class II region into mammalian cells by fusion with yeast spheroplasts carrying an artificial chromosome. J. Immun. 150, 5371–5378.PubMedGoogle Scholar
  15. 15.
    Featherstone, T. and Huxley, C. (1993) Extrachromosomal maintenance and amplification of yeast artificial chromosome DNA in mouse cells. Genomics 17, 267–278.PubMedCrossRefGoogle Scholar
  16. 16.
    Markie, D., Ragoussis, J., Senger, G., Rowan, A., Sansom, D., Trowsdale, J., et al. (1993) New vector for transfer of yeast artificial chromosomes to mammalian cells. Somat. Cell Mol. Genet. 19, 161–169.PubMedCrossRefGoogle Scholar
  17. 17.
    Nonet, G. H. and Wahl, G. M. (1993) Introduction of YACs containing a putative mammalian replication origin into mammalian cells can generate structures that replicate autonomously. Somat. Cell Mol. Genet. 19, 171–192.PubMedCrossRefGoogle Scholar
  18. 18.
    Silverman, G. A., Yang, E., Proffit, J. H., Zutter, M., and Korsmeyer, S. J. (1993) Genetic transfer and expression of reconstructed yeast artificial chromosomes containing normal and translocated BCL2 proto-oncogenes. Mol. Cell Biol. 13, 5469–5478.PubMedGoogle Scholar
  19. 19.
    Soh, J., Donnelly, R. J., Mariano, T. M., Cook, J. R., Schwartz, B., and Pestka, S. (1993) Identification of a yeast artificial chromosome clone encoding an accessory factor for the human interferon γ receptor: evidence for multiple accessory factors. Proc. Natl. Acad. Sci. USA 90, 8737–8741.PubMedCrossRefGoogle Scholar
  20. 20.
    Cook, J. R., Emanuel, S. L., Donnelly, R. J., Soh, J., Martano, T. M., Schwartz, B., et al. (1994) Sublocalization of the human interferon-γ receptor accessory factor gene and characterization of accessory factor activity by yeast artificial chromosomal fragmentation. J. Biol Chem. 269, 7013–7018.PubMedGoogle Scholar
  21. 21.
    Huxley, C. and Gnirke, A. (1991) Transfer of yeast artificial chromosomes from yeast to mammalian cells. BioEssays 13, 545–549.PubMedCrossRefGoogle Scholar
  22. 22.
    Wada, M., Ihara, Y., Tatsuka, M., Mitsui, H., Kohno, K., Kuwano, M., and Schlessinger, D. (1994) HPRT yeast artificial chromosome transfer into human cells by four methods and an involvement of homologous recombination. Biochem. Biophys. Res. Commun. 200, 1693–1700.PubMedCrossRefGoogle Scholar
  23. 23.
    Lamb, B. T., Sisodia, S. S., Lawler, A. M., Slunt, H. H., Kitt, C. A., Kearns, W. G., et al. (1993) Introduction and expression of the 400 kilobase precursor amyloid protein gene in transgenic mice. Nature Genet. 5, 22–30.PubMedCrossRefGoogle Scholar
  24. 24.
    Burgers, P. M. J. and Percival, K. J. (1987) Transformation of yeast spheroplasts without cell fusion. Anal. Biochem 163, 391–397.PubMedCrossRefGoogle Scholar
  25. 25.
    Eliceiri, B., Labella, T., Hagino, Y., Srivastava, A., Schlessinger, D., Pilia, G., et al. (1991) Stable integration and expression in mouse cells of yeast artificial chromosomes harboring human genes. Proc. Natl. Acad. Sci. USA 88, 2179–2183.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1996

Authors and Affiliations

  • Nicholas P. Davies
    • 1
  • Clare Huxley
    • 1
  1. 1.Department of Biochemistry and Molecular GeneticsSt. Mary’s Hospital Medical SchoolLondonUK

Personalised recommendations