Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells

Article metrics

  • An Erratum to this article was published on 05 September 1991

Abstract

GENE targeting in embryonic stem (ES) cells is a powerful tool for generating mice with null alleles1. Current methods of gene inactivation in ES cells introduce a neomycin gene (neo) cassette both as a mutagen and a selection marker for transfected cells2–11. Although null alleles are valuable, changes at the nucleotide level of a gene are very important for functional analysis. One gene family in which subtle mutations would be particularly valuable are the clusters of Hox homeobox genes12–16. Inactivation of genes in a cluster with a neo cassette that includes promoter/enhancer elements may deregulate transcription of neighbouring genes and generate a phenotype which is difficult to interpret. We describe here a highly efficient gene targeting method, termed the 'hit and run' procedure. This generates ES cells with subtle site-specific mutations with no selectable marker and may be useful for most genes. We have developed this procedure at the hypoxanthine phosphoribosyltransferase (hprt) locus and subsequently isolated ES cells with a premature stop codon in the homeobox of Hox-2.6 (ref. 14).

Access 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

    Capecchi, M. R. Science 244, 1288–1292 (1989).

  2. 2

    Schwartzberg, P. L., Goff, S. P. & Robertson, E. J. Science 246, 799–803 (1989).

  3. 3

    Zijlstra, M., Li, E., Sajjadi, F., Subramani, S. & Jaeniscu, R. Nature 342, 435–438 (1989).

  4. 4

    McMahon, A. P. & Bradley, A. Cell 62, 1073–1085 (1990).

  5. 5

    DeChiara, T. M., Efstratiadis, A. & Robertson, E. J. Nature 345, 78–80 (1990).

  6. 6

    Koller, B. H. & Smithies, O. Science 248, 1227–1230 (1990).

  7. 7

    Zijlstra, M. et al. Nature 344, 742–746 (1990).

  8. 8

    Thomas, K. R. & Capecchi, M. R. Cell 51, 503–512 (1987).

  9. 9

    Mansour, S. L., Thomas, K. R. & Capecchi, M. R. Nature 336, 348–352 (1988).

  10. 10

    Johnson, R. S. et al. Science 245, 1234–1236 (1989).

  11. 11

    Joyner, A. L., Skarnes, W. C. & Rossant, J. Nature 338, 153–156 (1989).

  12. 12

    Graham, A., Papalopulu, N. & Krumlauf, R. Cell 57, 367–378 (1989).

  13. 13

    Duboule, D. & Dolle, P. EMBO 8, 1497–1505 (1989).

  14. 14

    Graham, A. et al. Genes Dev. 2, 1424–1438 (1988).

  15. 15

    Gaunt, S. J., Krumlauf, R. & Doboule, D. Development 107, 131–142 (1989).

  16. 16

    Wright, C. V. E. et al. Trends biochem. Sci. 14, 52–56 (1989).

  17. 17

    Smithies, O., Gregg, R. G., Boggs, S. S., Koralewski, M. A. & Kucherlapati, R. S. Nature 317, 230–234 (1985).

  18. 18

    Doetschman, T. et al. Nature 330, 576–578 (1987).

  19. 19

    Baker, M. D. et al. Proc. natn. Acad. Sci. U.S.A. 85, 6432–6436 (1988).

  20. 20

    Baker, M. D., Pennell, N., Bosnoyan, L. & Schulman, M. J. Molec. cell. Biol. 9, 5500–5507 (1989).

  21. 21

    Borrelli, E., Heyman, R., Hsi, M. A. & Evans, R. M. Proc. natn. Acad. Sci. U.S.A. 85, 7572–7576 (1988).

  22. 22

    Melton, D. W., Konecki, D. S., Brennand, J. & Caskey, C. T. Proc. natn. Acad. Sci. U.S.A. 81, 2147–2151 (1984).

  23. 23

    Qian, Y. Q. et al. Cell 59, 573–580 (1989).

  24. 24

    Zimmer, A. & Gruss, P. Nature 338, 150–153 (1989).

  25. 25

    Holliday, R. Genet Res. 5, 282–304 (1964).

  26. 26

    Adra, C. N., Boer, P. H. & McBurney, M. W. Gene 60, 65–74 (1987).

  27. 27

    Pfarr, D. S. et al. DNA 5, 115–122 (1986).

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Hasty, P., Ramírez-Solis, R., Krumlauf, R. et al. Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells. Nature 350, 243–246 (1991) doi:10.1038/350243a0

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.