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The Blood-Brain Barrier pp 529–544Cite as

Mouse Genome Modification

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Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 89))

Abstract

The ability to alter the mouse genome through homologous recombination in their embryonic stem (ES) cells, and propagate the modification through their germ-line, has revolutionized biomedical research. Such gene-targeted mice have afforded researchers unprecedented opportunities to analyze gene function in vivo, and provided models for disease studies.

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References

  1. Evans, M. J., and Kaufman, M. H. (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature, 292, 154–156.

    Article  PubMed  CAS  Google Scholar 

  2. Martin, G. R. (1981) Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc. Natl. Acad. Sci. USA 78, 7634–7638.

    Article  PubMed  CAS  Google Scholar 

  3. Bradley, A., Evans, M., Kaufman, M. H., and Robertson, E. (1984) Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature 309, 255–256.

    Article  PubMed  CAS  Google Scholar 

  4. Smithies, O., Gregg, R. G., Boggs, S. S., Koralewski, M. A., and Kucherlapati, R. S. (1985) Insertion of DNA sequences into the human chromosomal beta-globin locus by homologous recombination. Nature 317, 230–234.

    Article  PubMed  CAS  Google Scholar 

  5. Thomas, K. R., and Capecchi, M. R. (1987) Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51, 503–512.

    Article  PubMed  CAS  Google Scholar 

  6. Doetschman, T., Maeda, N., and Smithies, O. (1988) Targeted mutation of the Hprt gene in mouse embryonic stem cells. Proc. Natl. Acad. Sci. USA 85, 8583–8587.

    Article  PubMed  CAS  Google Scholar 

  7. Koller, B. H., Hagemann, L. J., Doetschman, T., et al. (1989) Germ-line transmission of a planned alteration made in a hypoxanthine phosphoribosyltransferase gene by homologous recombination in embryonic stem cells. Proc. Natl. Acad. Sci. USA 86, 8927–8931.

    Article  PubMed  CAS  Google Scholar 

  8. Schwartzberg, P. L., Goff, S. P., and Robertson, E. J. (1989) Germ-line transmission of a c-abl mutation produced by targeted gene disruption in ES cells. Science 246, 799–803.

    Article  PubMed  CAS  Google Scholar 

  9. Thompson, L. H., and Schild, D. (1999) The contribution of homologous recombination in preserving genome integrity in mammalian cells. Biochimie 81, 87–105.

    Article  PubMed  CAS  Google Scholar 

  10. Hasty, P., Rivera-Perez, J., Chang, C., and Bradley, A. (1991) Target frequency and integration pattern for insertion and replacement vectors in embryonic stem cells. Mol. Cell Biol. 11, 4509–4517.

    PubMed  CAS  Google Scholar 

  11. Mansour, S. L., Thomas, K. R., and Capecchi, M. R. (1988) Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 336, 348–352.

    Article  PubMed  CAS  Google Scholar 

  12. Hasty, P., Abuin, A., and Bradley, A. (2000) Gene targeting, principles, and practice in mammalian cells. In Gene Targeting—A Practical Approach. 2 ed. Joyner, A. L., ed. Oxford University Press, New York, NY, pp. 1–35.

    Google Scholar 

  13. Lurquin, P. F. (1997) Gene transfer by electroporation. Mol. Biotechnol. 7, 5–35.

    Article  PubMed  CAS  Google Scholar 

  14. Andreason, G. L., and Evans, G. A. (1989) Optimization of electroporation for transfection of mammalian cell lines. Anal. Biochem. 180, 269–275.

    Article  PubMed  CAS  Google Scholar 

  15. Lupton, S. D., Brunton, L. L., Kalberg, V. A., and Overell, R. W. (1991) Dominant positive and negative selection using a hygromycin phosphotransferase-thymidine kinase fusion gene. Mol. Cell Biol. 11, 3374–3378.

    PubMed  CAS  Google Scholar 

  16. Chen, Y T., and Bradley, A. (2000) A new positive/negative selectable marker, puDeltatk, for use in embryonic stem cells. Genesis 28, 31–35.

    Article  PubMed  CAS  Google Scholar 

  17. Karreman, C. (1998) New positive/negative selectable markers for mammalian cells on the basis of Blasticidin deaminase-thymidine kinase fusions. Nucleic Acids Res. 26, 2508–2510.

    Article  PubMed  CAS  Google Scholar 

  18. Selfridge, J., Pow, A. M., McWhir, J., Magin, T. M., and Melton, D. W. (1992) Gene targeting using a mouse HPRT minigene/HPRT-deficient embryonic stem cell system: inactivation of the mouse ERCC-1 gene. Somat. Cell Mol. Genet. 18, 325–336.

    Article  PubMed  CAS  Google Scholar 

  19. Vasquez, K. M., Marburger, K., Intody, Z., and Wilson, J. H. (2001) Manipulating the mammalian genome by homologous recombination. Proc. Natl. Acad. Sci. USA 98, 8403–8410.

    Article  PubMed  CAS  Google Scholar 

  20. Yagi, T., Ikawa, Y., Yoshida, K., et al. (1990) Homologous recombination at c-fyn locus of mouse embryonic stem cells with use of diphtheria toxin A-fragment gene in negative selection. Proc. Natl. Acad. Sci. USA 87, 9918–9922.

    Article  PubMed  CAS  Google Scholar 

  21. Kobayashi, K., Ohye, T., Pastan, I., and Nagatsu, T. (1996) A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting. Nucleic Acids Res. 24, 3653–3655.

    Article  PubMed  CAS  Google Scholar 

  22. Donehower, L. A., Harvey, M., Slagle, B. L., et al. (1992) Mice deficient for p53 are devel-opmentally normal but susceptible to spontaneous tumours. Nature 356, 215–221.

    Article  PubMed  CAS  Google Scholar 

  23. Lindberg, R. L., Porcher, C., Grandchamp, B., et al. (1996) Porphobilinogen deaminase deficiency in mice causes a neuropathy resembling that of human hepatic porphyria. Nat. Genet. 12, 195–199.

    Article  PubMed  CAS  Google Scholar 

  24. te Riele, H., Maandag, E. R., and Berns, A. (1992) Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc. Natl. Acad. Sci. USA 89, 5128–5132.

    Article  Google Scholar 

  25. Mortensen, R. M., Conner, D. A., Chao, S., Geisterfer-Lowrance, A. A., and Seidman, J. G. (1992) Production of homozygous mutant ES cells with a single targeting construct. Mol. Cell Biol. 12, 2391–2395.

    PubMed  CAS  Google Scholar 

  26. Mortensen, R. M., Zubiaur, M., Neer, E. J., and Seidman, J. G (1991) Embryonic stem cells lacking a functional inhibitory G-protein subunit (alpha i2) produced by gene targeting of both alleles. Proc. Natl. Acad. Sci. USA 88, 7036–7040.

    Article  PubMed  CAS  Google Scholar 

  27. te Riele, H., Maandag, E. R., Clarke, A., Hooper, M., and Berns, A. (1990) Consecutive inactivation of both alleles of the pim-1 proto-oncogene by homologous recombination in embryonic stem cells. Nature 348, 649–651.

    Article  Google Scholar 

  28. Utomo, A. R., Nikitin, A. Y., and Lee, W. H. (1999) Temporal, spatial, and cell type-specific control of Cre-mediated DNA recombination in transgenic mice. Nat. Biotechnol. 17, 1091–1096.

    Article  PubMed  CAS  Google Scholar 

  29. Sauer, B. (1998) Inducible gene targeting in mice using the Cre/lox system. Methods 14, 381–392.

    Article  PubMed  CAS  Google Scholar 

  30. Torres, R. M., and Kuhn, R., eds. (1997) Laboratory protocols for conditional gene targeting. Oxford University Press, Oxford, UK.

    Google Scholar 

  31. Rajewsky, K., Gu, H., Kuhn, R., et al. (1996) Conditional gene targeting. J. Clin. Invest. 98, 600–603.

    Article  PubMed  CAS  Google Scholar 

  32. Wilson, T. J., and Kola, I. (2001) The LoxP/CRE system and genome modification. Methods Mol. Biol. 158, 83–94.

    PubMed  CAS  Google Scholar 

  33. Le, Y, and Sauer, B. (2000) Conditional gene knockout using cre recombinase. Methods Mol. Biol. 136, 477–485.

    PubMed  CAS  Google Scholar 

  34. Gossen, M., Freundlieb, S., Bender, G., Muller, G., Hillen, W., and Bujard, H. (1995) Transcriptional activation by tetracyclines in mammalian cells. Science 268, 1766–1769.

    Article  PubMed  CAS  Google Scholar 

  35. No, D., Yao, T P., and Evans, R. M. (1996) Ecdysone-inducible gene expression in mammalian cells and transgenic mice. Proc. Natl. Acad. Sci. USA 93, 3346–3351.

    Article  PubMed  CAS  Google Scholar 

  36. Danielian, P. S., Muccino, D., Rowitch, D. H., Michael, S. K., and McMahon, A. P. (1998) Modification of gene activity in mouse embryos in utero by a tamoxifen-inducible form of Cre recombinase. Curr. Biol. 8, 1323–1326.

    Article  PubMed  CAS  Google Scholar 

  37. Hayashi, S., and McMahon, A. P. (2002) Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse. Dev. Biol. 244, 305–318.

    Article  PubMed  CAS  Google Scholar 

  38. Dietrich, P., Dragatsis, I., Xuan, S., Zeitlin, S., and Efstratiadis, A. (2000) Conditional mutagenesis in mice with heat shock promoter-driven cre transgenes. Mamm. Genome 11, 196–205.

    Article  PubMed  CAS  Google Scholar 

  39. Gannon, M., Shiota, C., Postic, C., Wright, C. V., and Magnuson, M. (2000) Analysis of the Cre-mediated recombination driven by rat insulin promoter in embryonic and adult mouse pancreas. Genesis 26, 139–142.

    Article  PubMed  CAS  Google Scholar 

  40. Rossant, J., and McMahon, A. (1999) “Cre”-ating mouse mutants-a meeting review on conditional mouse genetics. Genes Dev. 13, 142–145.

    Article  PubMed  CAS  Google Scholar 

  41. Fiering, S., Epner, E., Robinson, K., et al. (1995) Targeted deletion of 5′HS2 of the murine beta-globin LCR reveals that it is not essential for proper regulation of the beta-globin locus. Genes Dev. 9, 2203–2213.

    Article  PubMed  CAS  Google Scholar 

  42. Valancius, V., and Smithies, O. (1991) Testing an “in-out” targeting procedure for making subtle genomic modifications in mouse embryonic stem cells. Mol. Cell Biol. 11, 1402–1408.

    PubMed  CAS  Google Scholar 

  43. Hasty, P., Ramirez-Solis, R., Krumlauf, R., and Bradley, A. (1991) Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells. Nature 350, 243–246.

    Article  PubMed  CAS  Google Scholar 

  44. Askew, G. R., Doetschman, T., and Lingrel, J. B. (1993) Site-directed point mutations in embryonic stem cells: a gene-targeting tag-and-exchange strategy. Mol. Cell Biol. 13, 4115–4124.

    PubMed  CAS  Google Scholar 

  45. Reid, L. H., Shesely, E. G., Kim, H. S., and Smithies, O. (1991) Cotransformation and gene targeting in mouse embryonic stem cells. Mol. Cell Biol. 11, 2769–2677.

    PubMed  CAS  Google Scholar 

  46. Davis, A. C., Wims, M., and Bradley, A. (1992) Investigation of coelectroporation as a method for introducing small mutations into embryonic stem cells. Mol. Cell Biol. 12, 2769–2776.

    PubMed  CAS  Google Scholar 

  47. Sambrook, J., Fritsch, E. F., and Maniatis, T (1989) Molecular Cloning—A Laboratory Manual. 2ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  48. Nagy, A., Rossant, J., Nagy, R., Abramow-Newerly, W., and Roder, J. C. (1993) Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc. Natl. Acad. Sci. USA 90, 8424–8428.

    Article  PubMed  CAS  Google Scholar 

  49. Smith, A. G., and Hooper, M. L. (1987) Buffalo rat liver cells produce a diffusible activity which inhibits the differentiation of murine embryonal carcinoma and embryonic stem cells. Dev. Biol. 121, 1–9.

    Article  PubMed  CAS  Google Scholar 

  50. Williams, R. L., Hilton, D. J., Pease, S., et al. (1988) Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336, 684–687.

    Article  PubMed  CAS  Google Scholar 

  51. Wood, S. A., Allen, N. D., Rossant, J., Auerbach, A., and Nagy, A. (1993) Non-injection methods for the production of embryonic stem cell-embryo chimaeras. Nature 365, 87–89.

    Article  PubMed  CAS  Google Scholar 

  52. Kawase, E., Suemori, H., Takahashi, N., Okazaki, K., Hashimoto, K., and Nakatsuji, N. (1994) Strain difference in establishment of mouse embryonic stem (ES) cell lines. Int. J. Dev. Biol. 38, 385–390.

    PubMed  CAS  Google Scholar 

  53. Doetschman, T., Gregg, R. G., Maeda, N., et al. (1987) Targeted correction of a mutant HPRT gene in mouse embryonic stem cells. Nature 330, 576–578.

    Article  PubMed  CAS  Google Scholar 

  54. Doetschman, T C., Eistetter, H., Katz, M., Schmidt, W., and Kemler, R. (1985) The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J. Embryol. Exp. Morphol. 87, 27–45.

    PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  56. Auerbach, W., Dunmore, J. H., Fairchild-Huntress, V., et al. (2000) Establishment and chimera analysis of 129/SvEv-and C57BL/6-derived mouse embryonic stem cell lines. Biotechniques 29, 1024–1028, 1030, 1032.

    PubMed  CAS  Google Scholar 

  57. Ledermann, B., and Burki, K. (1991) Establishment of a germ-line competent C57BL/6 embryonic stem cell line. Exp. Cell Res. 197, 254–258.

    Article  PubMed  CAS  Google Scholar 

  58. Noben-Trauth, N., Kohler, G., Burki, K., and Ledermann, B. (1996) Efficient targeting of the IL-4 gene in a BALB/c embryonic stem cell line. Transgenic Res. 5, 487–491.

    Article  PubMed  CAS  Google Scholar 

  59. Dinkel, A., Aicher, W.K., Warnatz, K., Burki, K., Eibel, H., and Ledermann, B. (1999) Efficient generation of transgenic BALB/c mice using BALB/c embryonic stem cells. J. Immunol. Methods 223, 255–260.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, University of Toronto, Toronto, Canada

    Richard Rozmahel

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  1. Richard Rozmahel
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Editors and Affiliations

  1. Division of Neuropathology, University of Toronto, Toronto, Canada

    Sukriti Nag

  2. Toronto Western Research Institute and Department of Pathology, University Health Network, Toronto, Canada

    Sukriti Nag

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© 2003 Humana Press Inc., Totowa, NJ

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Rozmahel, R. (2003). Mouse Genome Modification. In: Nag, S. (eds) The Blood-Brain Barrier. Methods in Molecular Medicine™, vol 89. Humana Press. https://doi.org/10.1385/1-59259-419-0:529

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  • DOI: https://doi.org/10.1385/1-59259-419-0:529

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-073-1

  • Online ISBN: 978-1-59259-419-1

  • eBook Packages: Springer Protocols

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