Skip to main content
Log in

Mouse genetics in the 21st century: using gene targeting to create a cornucopia of mouse mutants possessing precise genetic modifications

  • Invited Review Article
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Over 1500 mouse mutants have been identified, but few of the genes responsible for the defects have been identified. Recent developments in the area of gene targeting are revolutionizing the field of mouse genetics and our understanding of numerous genes, including those thought to be involved in cell proliferation and differentiation. Gene targeting was developed as a method for producing a predetermined mutation in a specific endogenous gene. Advances in the design of targeting vectors and in the use of embryonic stem cells have permitted the production of numerous mutant mice with null mutations in specific genes. These mutant mice will be critical for investigating thein vivo functions of many genes that have been cloned in recent years. This review discusses a wide range of new developments in the field of gene targeting with a focus on issues to be considered by those planning to use this new technology. It also examines some of the lessons learned from recent gene targeting studies and discusses different applications of the technology that are likely to generate scores of new animal models for a wide range of human diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

ES:

embryonic stem

neor :

neomycin resistance gene

HSV:

herpes simplex virus

tk:

thymidine kinase gene

PCR:

polymerase chain reaction

LIF:

leukemia inhibitory factor

LTP:

long-term potentiation

Rb:

retinoblastoma gene product

CF:

cystic fibrosis

References

  1. Alexander CM and Werb Z (1992) Targeted disruption of the tissue inhibitor of metalloproteinases gene increases the invasive behavior of primitive mesenchymal cells derived from embryonic stem cellsin vitro. J. Cell Biol. 118: 727–739.

    Google Scholar 

  2. Appleby MW, Gross JA, Cooke MP, Levin SD, Qian X and Perlmutter RM (1992) Defective T cell receptor signaling in mice lacking the thymic isoform of p59fyn. Cell 70: 751–763.

    Google Scholar 

  3. Axelrod HR and Lader (1983) A simplified method for obtaining embryonic stem cell lines from blastocysts. Cold Spring Harbor Conferences on Cell Proliferation 10: 665–670.

    Google Scholar 

  4. Baribault H and Oshima RH (1991) Polarized and functional epithelia can form after the targeted inactivation of both mouse keratin 8 alleles. J. Cell Biol. 115: 1675–1684.

    Google Scholar 

  5. Bradley A (1987) Production and analysis of chimaeric mice. In: Robertson EJ (ed.) Teratocarcinomas and Embryonic Stem Cells: A Practical Approach. IRL Press, Oxford, pp. 113–151.

    Google Scholar 

  6. Braun T, Rudnicki MA, Arnold H-H and Jaenisch R (1992) Targeted inactivation of the muscle regulatory geneMyf-5 results in results in abnormal rib development and perinatal death. Cell 71: 369–382.

    Google Scholar 

  7. Brinster RL, Braun RE, Lo D, Avarbock MR, Oram F and Palmitter RD (1989) Targeted correction of a major histocompatibility class II Eα gene by DNA microinjected into mouse eggs. Proc. Natl. Acad. Sci. USA 86: 7087–7091.

    Google Scholar 

  8. Capecchi MR (1989) The new mouse genetics: altering the genome by gene targeting. Trends Genet. 5: 70–76.

    Google Scholar 

  9. Charron J, Malynn BA, Robertson EJ, Goff SP and Alt FW (1990) High-frequency disruption of the N-myc gene in embryonic stem and pre-B cell lines by homologous recombination. Mol. Cell Biol. 10: 1799–1804.

    Google Scholar 

  10. Chisaka O and Capecchi MR (1991) Regionally restricted developmental defects resulting from targeted disruption of the mouse homeobox gene hox-1.5. Nature 350: 473–479.

    Google Scholar 

  11. Clarke AR, Maandag ER, van Roon M, van der Lugt NMT, van der Valk M, Hooper ML, Berns A and te Riele H (1992) Requirement for a functional Rb-1 gene in murine development. Nature 359: 328–330.

    Google Scholar 

  12. Clarke LL, Grubb BR, Gabriel SE, Smithies O, Koller BH and Boucher RC (1992) Defective epithelial chloride transport in a gene-targeted mouse model of cystic fibrosis. Science 257: 1125–1128.

    Google Scholar 

  13. Cosgrove D, Gray D, Dierich A, Kaufman J, Lemeur M, Benoist C and Mathis D (1991) Mice Lacking MHC class II molecules. Cell 66: 1051–1066.

    Google Scholar 

  14. DeChiara TM, Efstratiadis A and Robertson EJ (1990) A growth-deficiency phenotype in heterozygous mice carrying an insulin-like growth factor II gene disrupted by targeting. Nature 345: 78–80.

    Google Scholar 

  15. DeChiara TM, Robertson EJ and Efstratiadis A (1991) Parental imprinting of the mouse insulin-like growth factor II gene. Cell 64: 849–859.

    Google Scholar 

  16. Deng C and Capecchi MR (1992) Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus. Mol. Cell. Biol. 12: 3365–3371.

    Google Scholar 

  17. van Deursen J, Lovell-Badge R, Oerlemans F, Schepens J and Wieringa B (1991) Modulation of gene activity by consecutive gene targeting of one creatine kinase M allele in mouse embryonic stem cells. Nucl. Acids Res. 19: 2637–2643.

    Google Scholar 

  18. van Deursen J and Wieringa B (1992) Targeting of the creatine kinase M gene in embryonic stem cells using isogenic and nonisogenic vectors. Nucl. Acids Res. 20: 3815–3820.

    Google Scholar 

  19. Doetschman TC (1991) Gene targeting in embryonic stem cells. Biotechnology 16: 89–101.

    Google Scholar 

  20. Doetschman TC, Eistetter H, Katz M, Schmidt W and Kemler R (1985) Thein vitro development of blastocystderived embryonic stem cell lines: Formation of visceral yolk sac, blood islands and myocardium. J. Embryol. Exp. Morphol. 87: 27–45.

    Google Scholar 

  21. Donehower LA, Harvey M, Slagle BL, McArthur MJ, Montgomery CA Jr., Butel JS and Bradley A (1992) Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumors. Nature 356: 215–221.

    Google Scholar 

  22. Dorin JR, Dickinson P, Alton EWFW, Smith SN, Geddes DM, Stevenson NJ, Kimber WL, Fleming S, Clarke AR, Hooper ML, Anderson L, Beddington RSP and Porteous, DJ (1992) Cystic fibrosis in the mouse by targeted insertional mutagenesis. Nature 359: 211–215.

    Google Scholar 

  23. Evans MJ and Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292: 154–156.

    Google Scholar 

  24. Fung-Leung W-P and Mak TW (1992) Embryonic stem cells and homologous recombination. Current Opinion in Immunology 4: 189–194.

    Google Scholar 

  25. Fung-Leung W-P, Schilham MW, Rahemtulla A, Kundig TM, Vollenweider M, Potter J, van Ewijk W and Mak TW (1991) CD8 is needed for development of cytotoxic T cells but not helper T cells. Cell 65: 443–449.

    Google Scholar 

  26. Giese KP, Martini R, Lemke G, Soriano P and Schachner M (1992) Mouse Po gene disruption leads to hypomyelination, abnormal expression of recognition molecules, and degeneration of myelin and axons. Cell 71: 565–576.

    Google Scholar 

  27. Green MG (1989) Catalog of mutant genes and polymorphic loci. In: Lyon MF and Searle AG (eds.) Genetic Variants and Strains of the Laboratory Mouse. 2nd edn., Oxford University Press, Oxford, pp. 12–403.

    Google Scholar 

  28. Grusby MJ, Johnson RS, Papaioannou VE and Glimcher LH (1991) Depletion of CD4+ T cells in major histocompatibility complex II-deficient mice. Science 253: 1417–1420.

    Google Scholar 

  29. 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.

    Google Scholar 

  30. Herz J, Clouthier DE and Hammer RE (1992) LDL receptor-related protein internalizes and degrades uPA-PAI-1 complexes and is essential for embryo implantation. Cell 71: 411–421.

    Google Scholar 

  31. Hooper M, Hardy K, Handyside A, Hunter S and Monk M (1987) HPRT-deficient (Lesch-Nyhan) mouse embryos derived from germline colonization by cultured cells. Nature 326: 292–295.

    Google Scholar 

  32. Jacks T, Fazeli A, Schmitt EM, Bronson RT, Goodell MA and Weinberg RA (1992) Effects of an Rb mutation in the mouse. Nature 359: 295–300.

    Google Scholar 

  33. Jaenisch R (1988) Transgenic animals. Science 240: 1468–1474.

    Google Scholar 

  34. Jasin M and Berg P (1988) Homologous integration in mammalian cells without target gene selection. Genes Dev. 2: 1353–1361.

    Google Scholar 

  35. Jeannotte L, Ruiz JC and Robertson EJ (1991) Low level of Hox1.3 gene expression does not preclude the use of promoterless vectors to generate a targeted gene disruption. Mol. Cell. Biol. 11: 5578–5585.

    Google Scholar 

  36. Johnson RS, Sheng M, Greenberg ME, Kolodner RD, Papaioannou VE and Spiegelman BM (1989) Targeting of nonexpressed genes in embryonic stem cells via homologous recombination. Science 245: 1234–1236.

    Google Scholar 

  37. Johnson RS, Spiegelman BM and Papaioannou V (1992) Pleiotropic effects of a null mutation in the c-fos protooncogene. Cell 71: 577–586.

    Google Scholar 

  38. Joyner AL, Herrup K, Auerbach BA, Davis CA and Rossant J (1991) Subtle cerebellar phenotype in mice homozygous for a targeted deletion of the En-2 homeobox. Science 251: 1239–1243.

    Google Scholar 

  39. Joyner AL, Skarnes WC and Rossant, J (1989) Production of a mutation in mouse En-2 gene by homologous recombination in embryonic stem cells. Nature 338: 153–156.

    Google Scholar 

  40. Kim CG, Epner EM, Forrester WC and Groudine M (1992) Inactivation of the human β-globin gene by targeted insertion into the β-globin locus control region. Genes & Development 6: 928–938.

    Google Scholar 

  41. Kim H-S, Popovich BW, Shehee WR, Shesely EG and Smithies O (1991) Problems encountered in detecting a targeted gene by the polymerase chain reaction. Gene 103: 227–233.

    Google Scholar 

  42. Kim H-S and Smithies O (1988) Recombinant fragment assay for gene targeting based on the polymerase chain reaction. Nucl. Acids Res. 16: 8887–8930.

    Google Scholar 

  43. Kitamura D, Kudo A, Schaal S, Muller W, Melchers F and Rajewsky K (1992) A critical role of λ5 protein in B cell development. Cell 69: 823–831.

    Google Scholar 

  44. Kitamura D, Roes J, Kuhn R and Rajewsky K (1991) A B cell-deficient mouse by targeted disruption of the membrane exon of the immunoglobulin μ chain gene. Nature 350: 423–426.

    Google Scholar 

  45. Koller BH, Kim H-S, Latour AM, Brigman K, Boucher RC Jr., Scambler P, Wainwright B and Smithies O (1991) Toward an animal model of cystic fibrosis: targeted interruption of exon 10 of the cystic fibrosis transmembrane regulator gene in embryonic stem cells. Proc. Natl. Acad. Sci. USA 88: 10730–10734.

    Google Scholar 

  46. Koller BH, Marrack P, Kappler JW and Smithies O (1990) Normal development of mice deficient in β2M, MHCclass I protiens, and CD8+ T cells. Science 248: 1227–1230.

    Google Scholar 

  47. Koller BH and Smithies O (1989) Inactivating the β2-miscroglobulin locus in mouse embryonic stem cells by homologous recombination. Proc. Natl. Acad. Sci USA 86: 8932–8935.

    Google Scholar 

  48. Kuhn R, Rajewsky K and Muller W (1991) Generation and analysis of interleukin-4 deficient mice. Science 254: 707–710.

    Google Scholar 

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

    Google Scholar 

  50. Lee EY-HP, Chang C-Y, Hu N, Wang Y-CJ, Lai C-C, Herrup K, Lee W-H and Bradley A (1992) Mice deficient for Rb are nonviable and show defects in neurogenesis and haematopoiesis. Nature 359: 288–294.

    Google Scholar 

  51. Lee K-F, Li E, Huber LJ, Landis SC, Sharpe AH, Chao MV and Jaenisch R (1992) Targeted mutation of the gene encoding the low affinity NGF receptor p75 leads to deficits in the peripheral sensory nervous system. Cell 69: 737–749.

    Google Scholar 

  52. Li E, Bestor TH and Jaenisch R (1992) Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69: 915–926.

    Google Scholar 

  53. Liskay RM and Stachelek JL (1983) Evidence for intrachromosomal gene conversion in cultured mouse cells. Cell 35: 157–165.

    Google Scholar 

  54. Love PE, Tremblay ML and Westphal H (1992) Targeting of the T-cell receptor ζ-chain gene in embryonic stem cells: strategies for generating multiple mutations in a single gene. Proc. Natl. Acad. Sci. USA 89: 9929–9933.

    Google Scholar 

  55. Lovell-Badge RH (1987) Introduction of DNA into embryonic stem cells. In: Robertson, EJ (ed.) Teratocarcinomas and embryonic stem cells: a practical approach, IRL Press, Oxford, pp. 153–182.

    Google Scholar 

  56. Lufkin T, Dierich A, LeMeur M, Mark M and Chambon, P (1991) Disruption of the Hox-1.6 homeobox gene results in defects in a region corresponding to its rostral domain of expression. Cell 66: 1105–1119.

    Google Scholar 

  57. van der Lugt N, Maandag ER, te Riele H, Laird PW and Berns, A (1991) A pgk::hprt fusion as a selectable marker for targeting of genes in mouse embryonic stem cells: disruption of the T-cell receptor δ-chain-encoding gene. Gene 105: 263–267.

    Google Scholar 

  58. Magin TM, McWhir J and Melton DW (1992) A new mouse embryonic stem cell line with good germ line contribution and gene targeting frequency. Nucl. Acids Res. 20: 3795–3796.

    Google Scholar 

  59. Mansour SL, Thomas KR and Capecchi MR (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.

    Google Scholar 

  60. Mansour SL, Thomas KR, Deng C and Capecchi MR (1990) Introduction of a lacZ reporter gene into the mouse int-2 locus by homologous recombination. Proc. Natl. Acad. Sci. USA 87: 7688–7692.

    Google Scholar 

  61. Martin GR (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.

    Google Scholar 

  62. Martin GR and Lock LF (1983) Pluripotent cell lines derived from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Cold Spring Harbor Conferences on Cell Proliferation 10: 635–646.

    Google Scholar 

  63. McMahon AP and Bradley A (1990) The Wnt-1 (int-1) proto-oncogene in required for development of a large region of the mouse brain. Cell 62: 1073–1085.

    Google Scholar 

  64. Miller CCJ, McPheat JC and Potts WJ (1992) Targeted integration of the Ren-1D locus in mouse embryonic stem cells. Proc. Natl. Acad. Sci. USA 89: 5020–5024.

    Google Scholar 

  65. Moens CB, Auerbach AB, Conlon RA, Joyner AL and Rossant J (1992) A targeted mutation reveals a role for N-myc in branching morphogenesis in the embryonic mouse lung. Genes Dev. 6: 691–704.

    Google Scholar 

  66. Molina TJ, Kishihara K, Siderovski DP, van Ewijk W, Narendran A, Timms E, Wakeham A, Paige CJ, Hartmann K-U, Veillette A, Davidson D and Mak TW (1992) Profound block in thymocyte development in mice lacking p561ck. Nature 357: 161–164.

    Google Scholar 

  67. Mombaerts P, Clarke AR, hooper ML and Tonegawa S (1991) Creation of a large genomic deletion at the T-cell antigen receptor β-subunit locus in mouse embryonic stem cells by gene targeting. Proc. Natl. Acad. Sci. USA 88: 3084–3087.

    Google Scholar 

  68. Mombaerts P, Iacomini J, Johnson RS, Herrup K, Tonegawa S and Papaioannou VE (1992) RAG-1-deficient mice have no mature B and T lymphocytes. Cell 68: 869–877.

    Google Scholar 

  69. Mortensen RM, Conner DA, Chao S, Geisterfer-Lowrance AAT and Seidman JG (1992) Production of homologous mutant ES cells with a single targeting construct. Mol. Cell. Biol. 12: 2391–2395.

    Google Scholar 

  70. Mortensen RM, Zubiar M, Neer EJ and Seidman JG (1991) Embryonic stem cells lacking a functional inhibitory G-protein subunit (αi2) produced by gene targeting of both alleles. Proc. Natl. Acad. Sci. USA 88: 7036–7040.

    Google Scholar 

  71. Le Mouellic H, Lallemand Y and Brulet P (1992) Homeosis in the mouse induced by a null mutation in the Hox-3.1 gene. Cell 69: 251–264.

    Google Scholar 

  72. Le Mouellic H, Lallemand Y and Brulet P (1990) Targeted replacement of the homeobox gene Hox-3.1 by theEscherichia coli lacZ in mouse chimeric embryos. Proc. Natl. Acad. Sci. USA 87: 4712–4716.

    Google Scholar 

  73. Mucenski ML, McLain K Kier AB Swerdlow SH Schreiner CM, Miller TA, Pietryga DW, Scott WJ Jr. and Potter SS (1991) A functional c-myb gene is required for normal murine fetal hepatic hematopoiesis. Cell 65: 677–689.

    Google Scholar 

  74. Nagy A, Gocza E, Diaz EM, Prideaux VR, Ivanyi, E Markkula M and Rossant J (1990) Embryonic stem cells alone are able to support fetal development in the mouse. Devel. 110: 815–821.

    Google Scholar 

  75. Nichols J, Evans EP and Smith AG (1990) Establishment of germ-line-competent embryonic stem (ES) cells using differentiation inhibiting activity. Devel. 110: 1341–1348.

    Google Scholar 

  76. Pease S, Braghetta P, Gearing D, Grail D and Williams, RL (1990) Isolation of embryonic stem (ES) cells in media supplemented with recombinant leukemia inhibitory factor (LIF). Dev. Biol. 141: 344–352.

    Google Scholar 

  77. Pease S and Williams RL (1990) Formation of germ-line chimeras from embryonic stem cells maintained with recombinant leukemia inhibitory factor. Exp. Cell Res. 190: 209–211.

    Google Scholar 

  78. Pevny L, Simon MC, Robertson E, Klein WH, Tsai S-F, D'Agati V, Orkin SH and Constantini F (1991) Erythroid differentiation in chimaeric mice blocked by a targeted mutation in the gene for transcription factor GATA-1. Nature 349: 257–260.

    Google Scholar 

  79. Philpott KL, Viney JL, Kay G, Rastan S, Gardiner EM, Chae S, Hayday AC and Owen MJ (1992) Lymphoid development in mice congenitally lacking T cell receptor αβ-expressing cells. Science 256: 1448–1452.

    Google Scholar 

  80. Piedrahita JA, Zhang SH, Hagaman JR, Oliver PM and Maeda N (1992) Generation of mice carrying a mutant apolipoprotein E gene inactivated by gene targeting in embryonic stem cells. Proc. Natl. Acad. Sci. USA 89: 4471–4475.

    Google Scholar 

  81. Plump AS, Smith JD, Hayek T, Aalto-Sealto K, Walsh A, Verstuyft JG, Rubin EM and Breslow JL (1992) Severe hypercholesterolemia and atherosclerosis in apolipoprotein E-deficient mice created by homologous recombination in ES cells. Cell 71: 343–353.

    Google Scholar 

  82. Rahemtulla A, Fung-Leung WP, Schiiham MW, Kundig TM, Sambhara SR, Narendran A, Arabian A, Wakeham A, Paige CJ, Zinkernagel RM, Miller RG and Mak TW (1991) Normal development and function of CD8+ cells but markedly decreased helper cell activity in mice lacking CD4. Nature 353: 180–184.

    Google Scholar 

  83. Reid LH, Gregg RG, Smithies O and Koller BH (1990) Regulatory elements in the introns of the human HPRT gene are necessary for its expression in embryonic stem cells. Proc. Natl. Acad. Sci. USA 87: 4299–4303.

    Google Scholar 

  84. te Riele H, Maandag ER and Berns A (1992) Highly efficient gene recombination with isogenic DNA constructs. Proc. Natl. Acad. Sci USA 89: 5128–5132.

    Google Scholar 

  85. te Riele H, Maandag ER, 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.

    Google Scholar 

  86. Robertson EJ (1991) Using embryonic stem cells to introduce mutations into the mouse germ line. Biol. Reprod. 44: 238–245.

    Google Scholar 

  87. Robertson, EJ (1987) Embryo-derived stem cell lines. In: Robertson EJ (ed.) Teratocarcinomas and Embryonic Stem Cells. A Practical Approach, IRL Press, Oxford, pp. 71–112.

    Google Scholar 

  88. Robertson E, Bradley A, Kuehn M and Evans M (1986) Germ-line transmission of genes introduced into cultured pluripotential cells by retreviral vector. Nature 323: 445–448.

    Google Scholar 

  89. Robertson EJ, Kaufman MH, Bradley A and Evans MJ (1983) Isolation, properties, and karyotype analysis of pluripotential (EK) cell lines from normal and parthenogenetic embryos. Col Spring Harbor Conferences on Cell Proliferation 10: 647–663.

    Google Scholar 

  90. Roes J and Rajewsky K (1991) Cell autonomous expression of IgD is not essential for the maturation of conventional B cells. Int. Immunol. 3: 1367–1371.

    Google Scholar 

  91. Rudnicki MA, Braun T, Hinuma S and Jaenisch R (1992) Inactivation of MyoD in mice leads to up-regulation of the myogenic HLH geneMyf-5 and results in apparently normal muscle development. Cell 71: 383–390.

    Google Scholar 

  92. Saga Y, Yagi T, Ikawa Y, Sakakura T and Aizawa S (1992) Mice develop normally without tenascin. Genes & Development 6: 1821–1831.

    Google Scholar 

  93. Sawai S, Shimono A, Hanaoka K and Kondoh H (1991) Embryonic lethality resulting from disruption of both N-myc alleles in mouse zygotes. New Biol. 3: 861–869.

    Google Scholar 

  94. Schorle H, Holtschke T, Hunig T, Schimpl A and Horak I (1991) Development and function of T cells in mice rendered interleukin-2 deficient by gene targeting. Nature 352:621–624.

    Google Scholar 

  95. Schwartz F, Maeda N, Smithies O, Hickey R, Edelmann W, Skoultchi A and Kucherlapati R (1991) A dominant positive and negative selectable gene for use in mammalian cells. Proc. Natl. Acad. Sci. USA 88: 10416–10420.

    Google Scholar 

  96. Schwartzberg PL, Goff SP and Robertson EJ (1989) Germ-line transmission of a c-abl mutation produced by targeted gene disruption in ES cells. Science 246: 799–803.

    Google Scholar 

  97. Schwartzberg PL, Robertson EJ and Goff SP (1990) Targeted gene disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein. Proc. Natl. Acad. Sci. USA 87: 3210–3214.

    Google Scholar 

  98. Schwartzberg PL, Stall AM, Hardin JD, Bowdish KS, Humaran T, Boast S, Harbison ML, Robertson EJ and Goff SP (1991) Mice homozygous for the ablm1 mutation show poor viability and depletion of selected B and T cell populations. Cell: 65: 1165–1175.

    Google Scholar 

  99. Sedivy J and Joyner A (1992) Gene targeting. W.H. Freeman and Co., New York.

    Google Scholar 

  100. Shinkai Y, Tathbun G, Lam K-P, Oltz EM, Stewart V, Mendelsohn M, Charron J, Datta M, Young F, Stall AM and Alt FW (1992) RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell 68: 855–867.

    Google Scholar 

  101. Silva AJ, Stevens CF, Tonegawa S and Wang Y (1992a) Deficient hippocampal long-term potentiation in α-calcium-calmodulin kinase II mutant mice. Science 257: 201–206.

    Google Scholar 

  102. Silva AJ, Paylor R, Wehner JM and Tonegawa S (1992b) Impaired spatial learning in α-calcium-calmodulin kinase II mutant mice. Science 257: 206–211.

    Google Scholar 

  103. Smith AG and Hooper ML (1987) Buffalo rat liver cells produce a diffusible activity which inhibits the differentiation of murine embryonic carcinoma and embryonic stem cells. Dev. Biol. 121: 1–9.

    Google Scholar 

  104. Snouwaert JN, Brigman KK, Latour AM, Malouf NN, Boucher RC, Smithies O and Koller BH (1992) An animal model for cystic fibrosis made by gene targeting. Science 257: 1083–1088.

    Google Scholar 

  105. 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.

    Google Scholar 

  106. Stanton BR, Reid SW and Parada LF (1990) Germ line transmission of an inactive N-myc allele generated by homologous recombination in mouse embryonic stem cells. Mol. Cell. Biol. 10: 6755–6758.

    Google Scholar 

  107. Stein PL, Lee H-M, Rich S and Soriano P (1992) pp59fyn mutant mice display differential signaling in thymocytes and peripheral T cells. Cell 70: 741–750.

    Google Scholar 

  108. Stewart CL, Kasper P, Brunet LJ, Bhatt H, Gadi I, Kontgen F and Abbondanzo SJ (1992) Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor. Nature 359: 76–79.

    Google Scholar 

  109. Tarkowski AK, Witkowska A and Opas J (1977) Development of cytochalasin B-induced tetraploid and diploid/tetraploid mosaic mouse embryos. J. Embryol. Exp. Morph. 41: 47–64.

    Google Scholar 

  110. Thomas KR and Capecchi MR (1990) Targeted disruption of the murine int-1 proto-oncogene resulting in severe abnormalities in midorain and cerebellar development. Nature 346: 847–850.

    Google Scholar 

  111. Thomas KR and Capecchi MR (1987) Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell 51: 503–512.

    Google Scholar 

  112. Thomas KR and Capecchi MR (1986) Introduction of homologous DNA sequences into mammalian cells induces mutations in the cognate gene. Nature 324: 34–38.

    Google Scholar 

  113. Thomas KR, Deng C and Capecchi MR (1992) High-fiedelity gene targeting in embryonic stem cells by using sequence replacement vectors. Mol. Cell. Biol. 12: 2919–2923.

    Google Scholar 

  114. Thomas KR, Folger KR and Capecchi MR (1986) High frequency targeting of genes to specific sites in the mammalian genome. Cell 44: 419–428.

    Google Scholar 

  115. Travis J (1992) Scoring a technical knockout in mice. Science 256: 1392–1394.

    Google Scholar 

  116. Tybulewicz VLJ, Crawford CE, Jackson PK, Bronson RT and Mulligan RC (1991) Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene. Cell 65: 1153–1163.

    Google Scholar 

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

    Google Scholar 

  118. Valancius V and Smithies O (1991b) Double-strand gap repair in a mammalian gene targeting reaction. Mol. Cell. Biol. 11: 4389–4397

    Google Scholar 

  119. Ware LM and Axelrad AA (1972) Inherited resistance to N- and B-tropic murine leukemia virusesin vitro: evidence that congenic mouse strains SIM and SIM.R differ at the Fv-I locus. Virology 50: 339–348.

    Google Scholar 

  120. Williams RL, Hilton DJ, Pease S, Willson TA, Stewart CL Gearing, DP, Wagner EF, Metcalf D, Nicola NA and Gough NM (1988) Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336: 684–687.

    Google Scholar 

  121. Williamson R, Lee D, Hagman J and Maeda N (1992) Marked reduction of high density lipoprotein cholesterol in mice genetically modified to lack apolipoprotein A-I. Proc. Natl. Acad. Sci. USA 89: 7134–7138.

    Google Scholar 

  122. Yagi T, Ikawa Y, Yoshida K, Shigetani Y, Takeda N, Mabuchi I, Yamamoto T and Aizawa S (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.

    Google Scholar 

  123. Zijlstra M, Bix M, Simister NE, Loring JM, Raulet DH and Jaenisch R (1990) β2-microglobulin deficient mice lack CD4-8+ cytolytic T cells. Nature 344: 742–746.

    Google Scholar 

  124. Zijlstra M, Li E, Sajjadi F, Subramani S and Jaenisch R (1989) Germ-line transmission of a disrupted β2 gene produced by homologous recombination in embryonic stem cells. Nature 342: 435–438.

    Google Scholar 

  125. Zimmer A and Gruss P (1989) Production of chimaeric mice containing embryonic stem (ES) cells carrying a homeobox Hox 1.1 allele mutated by homologous recombination. Nature 338: 150–153.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wilder, P.J., Rizzino, A. Mouse genetics in the 21st century: using gene targeting to create a cornucopia of mouse mutants possessing precise genetic modifications. Cytotechnology 11, 79–99 (1993). https://doi.org/10.1007/BF00748997

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00748997

Key words

Navigation