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Transgenic mice containing a human heavy chain immunoglobulin gene fragment cloned in a yeast artificial chromosome

Nature Geneticsvolume 4pages117123 (1993) | Download Citation

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  • A Correction to this article was published on 01 July 1993

Abstract

We have developed a method for the introduction of yeast artificial chromosomes (YACs) into transgenic mice. An 85 kilobase (kb) fragment of the human heavy chain immunoglobulin gene was cloned as a YAC, and embryonic stem cell lines carrying intact, integrated YACs were derived by co–lipofection of the YAC with an unlinked selectable marker. Chimaeric founder animals were produced by blastocyst injection, and offspring transgenic for the YAC were obtained. Analysis of serum from these offspring for human heavy chain antibody subunits demonstrated expression of the YAC–borne immunoglobulin gene fragment. Co–lipofection may prove to be a highly–successful means of producing transgenic mice containing large gene fragments in YACs.

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References

  1. 1

    Jaenisch, R. Transgenic Animals. Science 240, 1468–1474 (1988).

  2. 2

    Brinster, R., Allen, J., Behringer, R., Gelinas, R. & Palmiter, R. Introns increase transcriptional efficiency in transgenic mice. Proc. natn. Acad. Sci. U.S.A. 85, 836–840 (1988).

  3. 3

    Burke, D., Carle, G. & Olson, M. Cloning of large segments of exogenous DNA into yeast artificial chromosomes. Science 236, 806–812 (1987).

  4. 4

    Pachnis, V., Pevney, L., Rothstein, R. & Costantini, F. Transfer of a yeast artificial chromosome carrying human DNA from Saccharomyces cerevisiae into human cells. Proc. natn. Acad. Sci. U.S.A. 87, 5109–5113 (1990).

  5. 5

    Pavan, W., Hieter, P. & Reeves, R. Modification and transfer into an embryonal carcinoma cell line of a 360-kilobase human-derived yeast artificial chromosome. Molec. cell Biol. 10, 4163–4169 (1990).

  6. 6

    Gnirke, A. et al. Cloning and in vivo expression of the human GART gene using yeast artificial chromosomes. EMBO J. 10, 1629–1634 (1991).

  7. 7

    Eliceiri, B. et al. Stable integration and expression in mouse cells of yeast artificial chromosomes harboring human genes. Proc. natn. Acad. Sci. U.S.A. 88, 2179–2183 (1991).

  8. 8

    Strauss, W. & Jaenisch, R. Molecular complementation of a collagen mutation in mammalian cells using yeast artificial chromosomes. EMBO J. 11, 417–422. (1992).

  9. 9

    Davies, N., Rosewell, I. & Bruggemann, M. Targeted alterations in yeast artificial chromosomes for inter-species gene transfer. Nucl. Acids Res. 20, 2693–2698 (1992).

  10. 10

    Schedl, A. et al. Transgenic mice generated by pronuclear injection of a yeast artificial chromosome. Nuc. Acids Res. 20, 3073–3077 (1992).

  11. 11

    Smith, D., Smyth, A. & Moir, D. Amplification of large artificial chomosomes. Proc. natn. Acad. Sci. U.S.A. 87, 8242–8246 (1990).

  12. 12

    Hofker, M., Walter, M. & Cox, D. Complete physical map of the human immunoglobulin heavy chain constant region gene complex. Proc. natn. Acad. Sci. U.S.A. 86, 5567–5571 (1989).

  13. 13

    Berman, J. et al. Content and organization of the human Ig VH locus: Definition of three new VH families and linkage to the Ig CH locus. EMBO J. 7, 727–738 (1988).

  14. 14

    Shin, E. et al. Physical map of the 3′ region of the human immunoglobulin heavy chain locus: clustering of autoantibody related variable segments in one haplotype. EMBO J. 10, 3641–3645 (1991).

  15. 15

    Traver, C.N., Klapholz, S., Hyman, R.W. & Davis, R.W. Rapid screening of a human genomic library in yeast artificial chromosomes for single-copy sequences. Proc. natn. Acad. Sci. U.S.A. 86, 5898–5902 (1989).

  16. 16

    Fox, T. et al. Analysis and manipulation of yeast mitochondrial genes, in Guide to Yeast Genetics and Molecular Biology (eds Guthrie, C. & Fink, G.) (Academic Press, San Diego, CA, 1991).

  17. 17

    Word, C., Blattner, F. & Kuzeil, W. The human immunoglobulin Cmu-Cdelta locus: complete nucleotide sequence and structural analysis. Int. Immunol. 1, 296–309 (1989).

  18. 18

    Ichihara, Y., Matsuoka, H. & Kurosawa, Y. Organization of human heavy chain diversity gene loci. EMBO J. 7, 4141–4150 (1988).

  19. 19

    Matsuda, F. et al. Dispersed localization of D segments in the human immunoglobulin heavy-chain locus. EMBO J. 7, 1047–1051 (1988).

  20. 20

    Soriano, P., Montgomery, C., Geske, R. & Bradley, A. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell 64, 693–702 (1991).

  21. 21

    Huxley, C., Hagino, Y., Schlessinger, D. & Olson, M. The human HPRT gene on a yeast artificial chromosome is functinal when transferred to mouse cells by cell fusion. Genomics 9, 742–750 (1991).

  22. 22

    Bradley, A. Production and analysis of chimaeric mice. in Teratocarcinomas and embryonic stem cells: a practical approach (ed. Robertson, E.) 113–152 (IRL Press, Oxford, 1987).

  23. 23

    Chen, J. et al. Immunoglobulin gene rearrangements in B cell deficient mice generated by targeted deletion of the JH locus. Int. Immun. (in the press).

  24. 24

    Pavan, W.J. & Reeves, R.H. Integrative selection of human chromosome-specific yeast artificial chromosomes. Proc. natn. Acad. Sci. U.S.A. 88, 7788–7791 (1991).

  25. 25

    Riley, J., Morten, J. & Anand, R. Targeted integration of neomycin into yeast artificial chromosomes (YACs) for transfection into mammalian cells. Nucl. Acids Res. 20, 2971–2976 (1992).

  26. 26

    de la Luna, S., Soria, I., Pulido, D., Ortin, J. & Jimenez, A. Efficient transformation of mammalian cells with constructs containing a puromycin-resistance marker. Gene 62, 121–126 (1988).

  27. 27

    Neil, D. et al. Structural Instability of human tandemly repeated DNA sequences cloned in yeast artificial chromosomal vectors. Nucl. Acids Res. 18, 1421–1428 (1990).

  28. 28

    Brown, S. XIST and the mapping of the X chromosome inactivation centre. BioEssays 13, 607–612 (1991).

  29. 29

    Gnirke, A. & Huxley, C. Transfer of the human HPRT and GART genes from yeast to mammalian cells by microinjection of YAC DNA. Somat. Cell molec. Genet. 17, 573–580 (1991).

  30. 30

    Ramirez-Solis, R., Davis, A.C. & Bradley, A. Gene targeting in mouse embryonic stem cells. Methods in Enzymology (in the press).

  31. 31

    Guise, J., Galli, G., Nevins, J. & Tucker, P. Developmental Regulation of Secreted and membrane Forms of Immunoglobulin μ chain. in Immunoglobulin Genes (eds Honjo, T., Alt, F. & Rabbitts, T.) 275–301 (Academic, San Diego, CA, 1989).

  32. 32

    Berman, J. et al. VH gene usage in humans: biased usage of the VH6 gene in immature B lymphoid cells. Eur. J. Immunol. 21, 1311–1314 (1991).

  33. 33

    Lubin, I. et al. Engranftment and development of human T and B cells in mice after bone marrow transplantation. Science 252, 427–431 (1991).

  34. 34

    Hawkins, R., Russell, S. & Winter, G. Selection of phage antibodies by binding affintiy: mimicking affinity maturation. J. molec. Biol. 226, 889–896 (1992).

  35. 35

    Strauss, W., Jaenisch, E. & Jaenisch, R. A strategy for rapid production and screening of yeast artificial chromosome libraries. Mamm. Genome 2, 150–157 (1992).

  36. 36

    Tybulewicz, V.L.J., Crawford, C.E., Jackson, P.J., Bronson, R.T. & Mulligan, R.C. Neonatal lethality and lympopenia in mice with a homozygous disruption of the c-abl proto-oncogene. Cell 40, 271–281 (1991).

  37. 37

    McMahon, A.P. & Bradley, A. The Wnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain. Cell 62, 1073–1085 (1990).

  38. 38

    Laird, P. et al. Simplified mammalian DNA isolation procedure. Nucl. Acids Res. 19, 4293 (1991).

  39. 39

    Ramirez-Solis, R. et al. Genomic DNA microextraction: a method to screen numerous samples. Anal. Biochem. 201, 331–335 (1992).

  40. 40

    Jakobovits, A. et al. Germ-line transmission and expression of a human-derived yeast artificial chromosome. Nature 362, 255–258 (1993).

  41. 41

    Schedl, A. et al. A yeast artificial chromosome covering the tyrosinase gene confers copy number-dependent expression in transgenic mice. Nature 362, 258–261 (1993).

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  1. GenPharm International, 297 N. Bernardo Avenue, Mountain View, California, 94043, USA

    • Ted K. Choi
    • , Paul W. Hollenbach
    • , Barbara E. Pearson
    • , Roanna M. Ueda
    • , Gregory N. Weddell
    • , Carole G. Kurahara
    • , Clive S. Woodhouse
    • , Robert M. Kay
    •  & Jeanne F. Loring

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https://doi.org/10.1038/ng0693-117

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