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Double Replacement Gene Targeting for the Production of a Series of Mouse Strains with Different Prion Protein Gene Alterations

Bio/Technology volume 13, pages 999–1004 (1995)Cite this article

Abstract

We have developed a double replacement gene targeting strategy which enables the production of a series of mouse strains bearing different subtle alterations to endogenous genes. This is a two-step process in which a region of the gene of interest is first replaced with a selectable marker to produce an inactivated allele, which is then re-targeted with a second vector to reconstruct the inactivated allele, concomitantly introducing an engineered mutation. Five independent embryonic stem cell lines have been produced bearing different targeted alterations to the prion protein gene, including one which raises the level of expression. We have constructed mice bearing the codon 101 proline to leucine substitution linked to the human familial prion disease, Gerstmann-Straussler-Scheinker syndrome. We anticipate that this procedure will have applications to the study of human inherited diseases and the development of therapies.

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References

  1. Melton, D.W. 1994. Gene targeting in the mouse. BioEssays 16: 633–638.

    Article  CAS  Google Scholar 

  2. 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  CAS  Google Scholar 

  3. Bradley, A., Evans, M., Kaufman, M.H. and Robertson, E. 1984. Formation of germline chimeras from embryo-derived teratocarcinoma cell lines. Nature 309: 255–256.

    Article  CAS  Google Scholar 

  4. Snouwaert, J.N., Brigman, K.K., Latour, A.M., Malouf, N.N., Boucher, R.C., Smithies, O. and Roller, B.H. 1992. An animal model for cystic fibrosis made by gene targeting. Science 257: 1083–1088.

    Article  CAS  Google Scholar 

  5. Dorin, J.R., Dickinson, P., Alton, E.W.F.W., Smith, S.N., Geddes, D.M., Stevenson, B.J., Kimber, W.L., Fleming, S., Clarke, A.R., Hooper, M.L., Anderson, L., Beddington, R.S.P. and Porteous, D.J. 1992. Cystic fibrosis in the mouse by targeted insemonal mutagenesis. Nature 359: 211–215.

    Article  CAS  Google Scholar 

  6. Tybulewicz, V.L.J., Tremblay, M.L., LaMarca, M.E., Willemsen, R., Stubblefleld, B.K., Winfield, S., Zablocka, B., Sidransky, E., Martin, B.M., Huang, S.P., Mintzer, K.A., Westphal, H., Mulligan, R.C. and Ginns, E.I. 1992. Animal model of Gaucher's disease from targeted disruption of the mouse glucocerebrosidase gene. Nature 357: 407–410.

    Article  CAS  Google Scholar 

  7. Rubin, E.M. and Smith, D.J. 1994. Atherosclerosis in mice: getting to the heart of a polygenic disorder. Trends in Genetics 10: 199–203.

    Article  CAS  Google Scholar 

  8. Kerem, B.-S., Rommens, J.M-., Buchanan, J.A., Markiewicz, D., Cox, T. K., Chakravarti, A., Buchwald, M. and Tsui, L.-C. 1989. Identification of a cystic fibrosis gene: genetic analysis. Science 245: 1073–1080.

    Article  CAS  Google Scholar 

  9. Prusiner, S.B. and Hsaio, K.K. 1994. Human prion diseases. Ann. Neurol. 35: 385–395.

    Article  CAS  Google Scholar 

  10. Prusiner, S.B. 1994. Inherited prion diseases. Proc. Natl. Acad. Sci. USA 91: 4611–4614.

    Article  CAS  Google Scholar 

  11. Palmer, M.S., Dryden, A.J., Hughes, T. and Collinge, J. 1991. Homozygous prion protein genotype predisposes to sporadic Creutzfeldt-Jakob disease. Nature 352: 340–341.

    Article  CAS  Google Scholar 

  12. Goldfarb, L.G., Petersen, R.B., Tabaton, M., Brown, P. et al. 1992. Fatal familial insomnia and familial Creutzfeldt-Jakob disease: disease phenotype determined by a DNA polymorphism. Science 258: 806–807.

    Article  CAS  Google Scholar 

  13. Carlson, G.A., Kingsbury, D.T., Goodman, P.A., Coleman, S., Marshall, S.T., DeArmond, S., Westaway, D. and Prusiner, S. B. 1986. Linkage of prion protein and scrapie incubation time genes. Cell 46: 503–511.

    Article  CAS  Google Scholar 

  14. Westaway, D., Goodman, P.A., Mirenda, C.A., McKinley, M.P., Carlson, G.A. and Prusiner, S.B. 1987. Distinct prion proteins in short and long scrapie incubation period mice. Cell 51: 651–662.

    Article  CAS  Google Scholar 

  15. 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  CAS  Google Scholar 

  16. Stacey, A., Schnieke, A., McWhir, J., Cooper, J., Colman, A. and Melton, D.W. 1994. Use of double-replacement gene targeting to replace the murine α-lactalbumin gene with its human counterpart in embryonic stem cells and mice. Mol. Cell. Biol. 14: 1009–1016.

    Article  CAS  Google Scholar 

  17. Magin, T.M., McWhir, J. and Melton, D.W. 1992. A new mouse embryonic stem cell line with good germ line contribution and gene targeting frequency. Nucl. Acid. Res. 20: 3795–3796.

    Article  CAS  Google Scholar 

  18. Westaway, D., Cooper, C., Turner, S., Da Costa, M., Carlson, G.A. and Prusiner, S.B. 1994. Structure and polymorphism of the mouse prion protein gene. Proc. Natl. Acad. Sci. USA 91: 6418–6422.

    Article  CAS  Google Scholar 

  19. Palmiter, R.D and Brinster, R.L. 1986. Germ-line transformation of mice. Ann. Rev. Genet. 20: 465–499.

    Article  CAS  Google Scholar 

  20. Scott, M., Foster, D., Mirenda, C., Serban, D., Coufal, F., Walchli, M., Torchia, M., Groth, D., Carlson, G., DeArmond, S.J., Westaway, D. and Prusiner, S.B. 1989. Transgenic mice expressing hamster prion protein produce species-specific scrapie infectivity and amyloid plaques. Cell 59: 847–857.

    Article  CAS  Google Scholar 

  21. Prusiner, S.B., Scott, M., Foster, D., Pan, K.-M., Groth, D., Mirenda, C., Torchia, M., Yang, S.-L., Serban, D., Carlson, G.A., Hoppe, P.C., Westaway, D. and DeArmond, S.J. 1990. Transgenetic studies implicate interactions between homologous PrP isoforms in scrapie prion replication. Cell 63: 673–686.

    Article  CAS  Google Scholar 

  22. Scott, M., Groth, D., Foster, D., Torchia, M., Yang, S.-L., DeArmond, S.J. and Prusiner, S.B. 1993. Propagation of prions with artificial properties in transgenic mice expressing chimeric PrP genes. Cell 73: 979–988.

    Article  CAS  Google Scholar 

  23. Hsaio, K.K., Scott, M., Foster, D., Groth, D.F., DeArmond, S.J. and Prusiner, S.B. 1990. Spontaneous neurodegeneration in transgenic mice with mutant prion protein. Science 250: 1587–1590.

    Article  Google Scholar 

  24. Westaway, D., DeArmond, S.J., Cayetano-Canlas, J., Groth, D., Foster, D., Yang, S.-L., Torchia, M., Carlson, G.A. and Prusiner, S.B. 1994. Degeneration of skeletal muscie, peripheral nerves, and the central nervous system in transgenic mice overexpressing wild- type prioh proteins. Cell 76: 117–129.

    Article  CAS  Google Scholar 

  25. Games, D., Adams, D., Alessandrini, R., Barbour, R. et al. 1995. Alzheimer-type neuropathology in transgenic mice overexpressing V717F α-amyloid precursor protein. Nature 373: 523–527.

    Article  CAS  Google Scholar 

  26. Xu, Z., Cork, L.C., Griffin, J.W. and Cleveland, D.W. 1993. Increased expression of neurofilament subunit NF-L produces morphological alterations that resemble the pathology of human motor neuron disease. Cell 73: 23–33.

    Article  CAS  Google Scholar 

  27. Cote, F., Collard, J.-F. and Julien, J.-P. 1993. Progressive neuronopathy in transgenic mice expressing the human neurofilament heavy gene: a mouse model for amylotrophic lateral sclerosis. Cell 73: 35–46.

    Article  CAS  Google Scholar 

  28. Deng, C., Thomas, K.T. and Capecchi, M.R. 1993. Location of crossovers during gene targeting with insertion and replacement vectors. Mol. Cell. Biol. 13: 2134–2140.

    Article  CAS  Google Scholar 

  29. Taki, S., Meiering, M. and Rajewsky, K. 1993. Targeted insertion of a variable region gene into the immunoglobulin heavy chain locus. Science 262: 1268–1271.

    Article  CAS  Google Scholar 

  30. Detloff, P.J., Lewis, J., John, S.W., Shehee, W.R., Langenbach, R., Maeda, N. and Smithies, O. 1994. Deletion and replacement of the mouse adult β-globin genes by a “plug and socket” repeated targeting strategy. Mol. Cell. Biol. 14: 6936–6943.

    Article  CAS  Google Scholar 

  31. Valancius, V. and Smithies, O. 1991. Testing an “in-out” targeting procedure for making subtle alterations in mouse embryonic stem cells. Mol. Cell. Biol. 11: 1402–1408.

    Article  CAS  Google Scholar 

  32. Ramirez-Solis, R., Zeng, H., Whiting, J., Krumlauf, R. and Bradley, A. 1993. Hoxb-4 {Hox 2.6) mutant mice show homeotic transformation of a cervical vertebra and defects in the closure of the sternal rudiments. Cell 73: 279–294.

    Article  CAS  Google Scholar 

  33. Wu, H., Liu, X. and Jaenisch, R. 1994. Double replacement: strategy for efficient introduction of subtle mutations into the murine Collα-1 gene by homologous recombination irk embryonic stem cells. Proc. Natl. Acad. Sci. USA 91: 2819–2823.

    Article  CAS  Google Scholar 

  34. Ernst, M., Gearing, D.P. and Dunn, A.R. 1994. Functional and biochemical association of Hck with the LlF/IL-6 receptor signal transducing subunit gpl30 in embryonic stem cells. EMBO J. 13: 1574–1584.

    Article  CAS  Google Scholar 

  35. Reid, L.H., Gregg, R.G., Smithies, O. and Koller, B.H. 1990. Regulatory elements in the introns of the humart HPRT gene are neccessary for its expression in embryonic stem cells. Proc. Natl. Acad. Sci. USA 87: 4299–4303.

    Article  CAS  Google Scholar 

  36. 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.

    Article  CAS  Google Scholar 

  37. Kunkel, T.A., Roberts, J.D. and Zakour, R.A. 1987. Rapid and efficient site specific mutagenesis without phenotypic selection. Meth. Enzymol. 154: 367–382.

    Article  CAS  Google Scholar 

  38. Magin, T.M., McEwan, C., Milne, M., Pow, A.M., Selfridge, J. and Melton, D.W. 1992. A position- and orientation-dependent element in the first intron is required for expression of the mouse hprt gene in embryonic stem cells. Gene 122: 289–296.

    Article  CAS  Google Scholar 

  39. 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. Som. Cell. Mol. Genet. 18: 325–336.

    Article  CAS  Google Scholar 

  40. Miller, S.A., Dykes, D.D. and Polesky, H.F. 1988. A simple salting out procedure for extracting DNA from human nucleated cells. Nucl. Acid. Res. 16: 1215.

    Article  CAS  Google Scholar 

  41. Thompson, S., Clarke, A.R., Pow, A.M., Hooper, M.L. and Melton, D.W. 1989. Germ line transmission and expression of a corrected HPRT gene produced by gene targeting in embryonic stem cells. Cell 56: 313–321.

    Article  CAS  Google Scholar 

  42. Melton, D.W., Konecki, D.S., Ledbetter, D.H., Hejtmancik, J.F. and Caskey, C.T. 1981. In vitro translation of hypoxanthine/guanine phosphoribosyltrans-ferase mRNA; characterisation of a mouse neuroblastoma cell line that has elevated levels of hypoxanthine/guanine phosphoribosyltransferase protein. Proc. Natl. Acad. Sci. USA 78: 6977–6980.

    Article  CAS  Google Scholar 

  43. Minty, A.J., Caravatti, M., Robert, B., Cohen, A., Daubas, P., Weydert, A., Gros, F. and Buckingham, M. 1981. Mouse actin messenger RNAs: construction and characterization of a recombinant plasmid molecule containing a complementary DNA transcript of mouse α-actin mRNA. J. Biol. Chem. 256: 1008–1014.

    CAS  Google Scholar 

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Authors and Affiliations

  1. Institute of Cell and Molecular Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh, EH9 3JR, Scotland

    Richard C. Moore, Nicola J. Redhead, Jim Selfridge & David W. Melton

  2. BBSRC and MRC Neuropathogenesis Unit, Institute for Animal Health, West Mains Road, Edinburgh, EH9 3JF, Scotland

    James Hope & Jean C. Manson

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  1. Richard C. Moore
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Correspondence to David W. Melton.

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Moore, R., Redhead, N., Selfridge, J. et al. Double Replacement Gene Targeting for the Production of a Series of Mouse Strains with Different Prion Protein Gene Alterations. Nat Biotechnol 13, 999–1004 (1995). https://doi.org/10.1038/nbt0995-999

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