Generating Conditional Knockout Mice

Methods in Molecular Biology

Volumn 693, Issue , 2011, Pages 205-231

  Friedel, Roland H ^a   Wurst, Wolfgang ^b,c   Wefers, Benedikt ^b   Kühn, Ralf ^b  

^a MOUNT SINAI SCHOOL OF MEDICINE   (United States)

^b INSTITUTE OF EPIDEMIOLOGY   (Germany)

^c MAX PLANCK INSTITUTE OF PSYCHIATRY   (Germany)

Author keywords

Conditional mutant; Cre; ES cells; Inducible; loxP; Mouse mutant; Tamoxifen

Indexed keywords

CRE RECOMBINASE; INTEGRASE; TAMOXIFEN;

ANIMAL; ARTICLE; CYTOLOGY; DRUG EFFECT; EMBRYONIC STEM CELL; GENE EXPRESSION REGULATION; GENETICS; METABOLISM; MOUSE; MOUSE MUTANT; MUTAGENESIS; TRANSGENIC MOUSE;

ANIMALS; EMBRYONIC STEM CELLS; GENE EXPRESSION REGULATION; INTEGRASES; MICE; MICE, KNOCKOUT; MICE, TRANSGENIC; MUTAGENESIS; TAMOXIFEN;

MLCS; MLOWN;

EID: 79952276543     PISSN: 10643745     EISSN: 19406029     Source Type: Book Series    
DOI: 10.1007/978-1-60761-974-1_12     Document Type: Chapter

References (90)

1. Capecchi, M. R. (1989) The new mouse genetics: altering the genome by gene targeting, Trends Genet 5, 70–76.
2. Mak, T. (1998) The Gene Knockout Facts Book, Academic Press, San Diego.
3. Gu, H., Marth, J. D., Orban, P. C., Mossmann, H., and Rajewsky, K. (1994) Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting, Science 265, 103–106.
4. Rajewsky, K., Gu, H., Kuhn, R., Betz, U. A., Muller, W., Roes, J., and Schwenk, F. (1996) Conditional gene targeting, J Clin Invest 98, 600–603.
5. Branda, C. S. and Dymecki, S. M. (2004) Talking about a revolution: the impact of site-specific recombinases on genetic analyses in mice, Dev Cell 6, 7–28.
6. Sauer, B. and McDermott, J. (2004) DNA recombination with a heterospecific Cre homolog identified from comparison of the pac-c1 regions of P1-related phages, Nucleic Acids Res 32, 6086–6095.
7. Anastassiadis, K., Fu, J., Patsch, C., Hu, S., Weidlich, S., Duerschke, K., Buchholz, F., Edenhofer, F., and Stewart, A. F. (2009) Dre recombinase, like Cre, is a highly efficient site-specific recombinase in E. coli, mammalian cells and mice, Dis Model Mech 2, 508–515.
8. Hoess, R. H. and Abremski, K. (1984) Interaction of the bacteriophage P1 recombinase Cre with the recombining site loxP, Proc Natl Acad Sci U S A 81, 1026–1029.
9. Abremski, K., Hoess, R., and Sternberg, N. (1983) Studies on the properties of P1 site-specific recombination: evidence for topologically unlinked products following recombination, Cell 32, 1301–1311.
10. Hoess, R. H., Wierzbicki, A., and Abremski, K. (1986) The role of the loxP spacer region in P1 site-specific recombination, Nucleic Acids Res 14, 2287–2300.
11. Indra, A. K., Warot, X., Brocard, J., Bornert, J. M., Xiao, J. H., Chambon, P., and Metzger, D. (1999) Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases, Nucleic Acids Res 27, 4324–4327.
12. 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 U S A 89, 5128–5132.
13. Kontgen, F., Suss, G., Stewart, C., Steinmetz, M., and Bluethmann, H. (1993) Targeted disruption of the MHC class II Aa gene in C57BL/6 mice, Int Immunol 5, 957–964.
14. Pettitt, S. J., Liang, Q., Rairdan, X. Y., Moran, J. L., Prosser, H. M., Beier, D. R., Lloyd, K. C., Bradley, A., and Skarnes, W. C. (2009) Agouti C57BL/6 N embryonic stem cells for mouse genetic resources, Nat Methods 6, 493–495.
15. Hitz, C., Wurst, W., and Kuhn, R. (2007) Conditional brain-specific knockdown of MAPK using Cre/loxP regulated RNA interference, Nucleic Acids Res 35, e90.
16. Eggan, K., Akutsu, H., Loring, J., Jackson-Grusby, L., Klemm, M., Rideout, W. M., 3rd, Yanagimachi, R., and Jaenisch, R. (2001) Hybrid vigor, fetal overgrowth, and viability of mice derived by nuclear cloning and tetraploid embryo complementation, Proc Natl Acad Sci U S A 98, 6209–6214.
17. Southon, E. and Tessarollo, L. (2009) Manipulating mouse embryonic stem cells, Methods Mol Biol 530, 165–185.
18. Rodriguez, C. I., Buchholz, F., Galloway, J., Sequerra, R., Kasper, J., Ayala, R., Stewart, A. F., and Dymecki, S. M. (2000) High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP, Nat Genet 25, 139–140.
19. Testa, G., Schaft, J., van der Hoeven, F., Glaser, S., Anastassiadis, K., Zhang, Y., Hermann, T., Stremmel, W., and Stewart, A. F. (2004) A reliable lacZ expression reporter cassette for multipurpose, knockout-first alleles, Genesis 38, 151–158.
20. Schnutgen, F., De-Zolt, S., Van Sloun, P., Hollatz, M., Floss, T., Hansen, J., Altschmied, J., Seisenberger, C., Ghyselinck, N. B., Ruiz, P., Chambon, P., Wurst, W., and von Melchner, H. (2005) Genomewide production of multipurpose alleles for the functional analysis of the mouse genome, Proc Natl Acad Sci U S A 102, 7221–7226.
21. Choi, T., Huang, M., Gorman, C., and Jaenisch, R. (1991) A generic intron increases gene expression in transgenic mice, Mol Cell Biol 11, 3070–3074.
22. Pfarr, D. S., Rieser, L. A., Woychik, R. P., Rottman, F. M., Rosenberg, M., and Reff, M. E. (1986) Differential effects of polyadenylation regions on gene expression in mammalian cells, DNA 5, 115–122.
23. Le, Y., Gagneten, S., Tombaccini, D., Bethke, B., and Sauer, B. (1999) Nuclear targeting determinants of the phage P1 cre DNA recombinase, Nucleic Acids Res 27, 4703–4709.
24. Shimshek, D. R., Kim, J., Hubner, M. R., Spergel, D. J., Buchholz, F., Casanova, E., Stewart, A. F., Seeburg, P. H., and Sprengel, R. (2002) Codon-improved Cre recombinase (iCre) expression in the mouse, Genesis 32, 19–26.
25. Parkitna, J. R., Engblom, D., and Schutz, G. (2009) Generation of Cre recombinase-expressing transgenic mice using bacterial artificial chromosomes, Methods Mol Biol 530, 325–342.
26. Hentze, M. W. and Kulozik, A. E. (1999) A perfect message: RNA surveillance and non-sense-mediated decay, Cell 96, 307–310.
27. Olson, E. N., Arnold, H. H., Rigby, P. W., and Wold, B. J. (1996) Know your neighbors: three phenotypes in null mutants of the myogenic bHLH gene MRF4, Cell 85, 1–4.
28. Lindeberg, J. and Ebendal, T. (1999) Use of an internal ribosome entry site for bicistronic expression of Cre recombinase or rtTA transactivator, Nucleic Acids Res 27, 1552–1554.
29. Gorski, J. A. and Jones, K. R. (1999) Efficient bicistronic expression of cre in mammalian cells, Nucleic Acids Res 27, 2059–2061.
30. Szymczak, A. L., Workman, C. J., Wang, Y., Vignali, K. M., Dilioglou, S., Vanin, E. F., and Vignali, D. A. (2004) Correction of multigene deficiency in vivo using a single “self-cleaving” 2A peptide-based retroviral vector, Nat Biotechnol 22, 589–594.
31. Nagy, A., Mar, L., and Watts, G. (2009) Creation and use of a cre recombinase transgenic database, Methods Mol Biol 530, 365–378.
32. Kuhn, R., Schwenk, F., Aguet, M., and Rajewsky, K. (1995) Inducible gene targeting in mice, Science 269, 1427–1429.
33. Saam, J. R. and Gordon, J. I. (1999) Inducible gene knockouts in the small intestinal and colonic epithelium, J Biol Chem 274, 38071–38082.
34. 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.
35. Brooks, A. I., Muhkerjee, B., Panahian, N., Cory-Slechta, D., and Federoff, H. J. (1997) Nerve growth factor somatic mosaicism produced by herpes virus-directed expression of cre recombinase, Nat Biotechnol 15, 57–62.
36. Anton, M. and Graham, F. L. (1995) Site-specific recombination mediated by an adenovirus vector expressing the Cre recombinase protein: a molecular switch for control of gene expression, J Virol 69, 4600–4606.
37. Kaspar, B. K., Vissel, B., Bengoechea, T., Crone, S., Randolph-Moore, L., Muller, R., Brandon, E. P., Schaffer, D., Verma, I. M., Lee, K. F., Heinemann, S. F., and Gage, F. H. (2002) Adeno-associated virus effectively mediates conditional gene modification in the brain, Proc Natl Acad Sci U S A 99, 2320–2325.
38. Indra, A. K., Li, M., Brocard, J., Warot, X., Bornert, J. M., Gerard, C., Messaddeq, N., Chambon, P., and Metzger, D. (2000) Targeted somatic mutagenesis in mouse epidermis, Horm Res 54, 296–300.
39. Feil, S., Valtcheva, N., and Feil, R. (2009) Inducible Cre mice, Methods Mol Biol 530, 343–363.
40. Schmidt-Supprian, M. and Rajewsky, K. (2007) Vagaries of conditional gene targeting, Nat Immunol 8, 665–668.
41. Vogt, M. A., Chourbaji, S., Brandwein, C., Dormann, C., Sprengel, R., and Gass, P. (2008) Suitability of tamoxifen-induced muta- genesis for behavioral phenotyping, Exp Neurol 211, 25–33.
42. Erdmann, G., Schutz, G., and Berger, S. (2007) Inducible gene inactivation in neurons of the adult mouse forebrain, BMC Neurosci 8, 63.
43. Kiermayer, C., Conrad, M., Schneider, M., Schmidt, J., and Brielmeier, M. (2007) Optimization of spatiotemporal gene inactivation in mouse heart by oral application of tamoxifen citrate, Genesis 45, 11–16.
44. Bruning, J. C., Michael, M. D., Winnay, J. N., Hayashi, T., Horsch, D., Accili, D., Goodyear, L. J., and Kahn, C. R. (1998) A muscle-specific insulin receptor knockout exhibits features of the metabolic syndrome of NIDDM without altering glucose tolerance, Mol Cell 2, 559–569.
45. Kulkarni, R. N., Bruning, J. C., Winnay, J. N., Postic, C., Magnuson, M. A., and Kahn, C. R. (1999) Tissue-specific knockout of the insulin receptor in pancreatic beta cells creates an insulin secretory defect similar to that in type 2 diabetes, Cell 96, 329–339.
46. Michael, M. D., Kulkarni, R. N., Postic, C., Previs, S. F., Shulman, G. I., Magnuson, M. A., and Kahn, C. R. (2000) Loss of insulin signaling in hepatocytes leads to severe insulin resistance and progressive hepatic dysfunction, Mol Cell 6, 87–97.
47. Kwan, K. M. (2002) Conditional alleles in mice: practical considerations for tissue-specific knockouts, Genesis 32, 49–62.
48. Schwenk, F., Baron, U., and Rajewsky, K. (1995) A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells, Nucleic Acids Res 23, 5080–5081.
49. Minichiello, L., Korte, M., Wolfer, D., Kuhn, R., Unsicker, K., Cestari, V., Rossi-Arnaud, C., Lipp, H. P., Bonhoeffer, T., and Klein, R. (1999) Essential role for TrkB receptors in hippocampus-mediated learning, Neuron 24, 401–414.
50. Tronche, F., Kellendonk, C., Kretz, O., Gass, P., Anlag, K., Orban, P. C., Bock, R., Klein, R., and Schutz, G. (1999) Disruption of the glucocorticoid receptor gene in the nervous system results in reduced anxiety, Nat Genet 23, 99–103.
51. Soriano, P. (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain, Nat Genet 21, 70–71.
52. Mao, X., Fujiwara, Y., and Orkin, S. H. (1999) Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice, Proc Natl Acad Sci U S A 96, 5037–5042.
53. Kawamoto, S., Niwa, H., Tashiro, F., Sano, S., Kondoh, G., Takeda, J., Tabayashi, K., and Miyazaki, J. (2000) A novel reporter mouse strain that expresses enhanced green fluorescent protein upon Cre-mediated recombination, FEBS Lett 470, 263–268.
54. Srinivas, S., Watanabe, T., Lin, C. S., William, C. M., Tanabe, Y., Jessell, T. M., and Costantini, F. (2001) Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus, BMC Dev Biol 1, 4.
55. Luche, H., Weber, O., Nageswara Rao, T., Blum, C., and Fehling, H. J. (2007) Faithful activation of an extra-bright red fluorescent protein in “knock-in” Cre-reporter mice ideally suited for lineage tracing studies, Eur J Immunol 37, 43–53.
56. Muzumdar, M. D., Tasic, B., Miyamichi, K., Li, L., and Luo, L. (2007) A global double-fluorescent Cre reporter mouse, Genesis 45, 593–605.
57. Lobe, C. G., Koop, K. E., Kreppner, W., Lomeli, H., Gertsenstein, M., and Nagy, A. (1999) Z/AP, a double reporter for cre-mediated recombination, Dev Biol 208, 281–292.
58. Novak, A., Guo, C., Yang, W., Nagy, A., and Lobe, C. G. (2000) Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cre-mediated excision, Genesis 28, 147–155.
59. Seibler, J., Zevnik, B., Kuter-Luks, B., Andreas, S., Kern, H., Hennek, T., Rode, A., Heimann, C., Faust, N., Kauselmann, G., Schoor, M., Jaenisch, R., Rajewsky, K., Kuhn, R., and Schwenk, F. (2003) Rapid generation of inducible mouse mutants, Nucleic Acids Res 31, e12.
60. Hameyer, D., Loonstra, A., Eshkind, L., Schmitt, S., Antunes, C., Groen, A., Bindels, E., Jonkers, J., Krimpenfort, P., Meuwissen, R., Rijswijk, L., Bex, A., Berns, A., and Bockamp, E. (2007) Toxicity of ligand-dependent Cre recombinases and generation of a conditional Cre deleter mouse allowing mosaic recombination in peripheral tissues, Physiol Genomics 31, 32–41.
61. Santagati, F., Minoux, M., Ren, S. Y., and Rijli, F. M. (2005) Temporal requirement of Hoxa2 in cranial neural crest skeletal morphogenesis, Development 132, 4927–4936.
62. 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.
63. Gruber, M., Hu, C. J., Johnson, R. S., Brown, E. J., Keith, B., and Simon, M. C. (2007) Acute postnatal ablation of Hif-2alpha results in anemia, Proc Natl Acad Sci U S A 104, 2301–2306.
64. Li, M., Indra, A. K., Warot, X., Brocard, J., Messaddeq, N., Kato, S., Metzger, D., and Chambon, P. (2000) Skin abnormalities generated by temporally controlled RXRalpha mutations in mouse epidermis, Nature 407, 633–636.
65. Yajima, I., Belloir, E., Bourgeois, Y., Kumasaka, M., Delmas, V., and Larue, L. (2006) Spatiotemporal gene control by the Cre-ERT2 system in melanocytes, Genesis 44, 34–43.
66. Bosenberg, M., Muthusamy, V., Curley, D. P., Wang, Z., Hobbs, C., Nelson, B., Nogueira, C., Horner, J. W., II, Depinho, R., and Chin, L. (2006) Characterization of melanocyte-specific inducible Cre recombinase transgenic mice, Genesis 44, 262–267.
67. Imai, T., Chambon, P., and Metzger, D. (2000) Inducible site-specific somatic mutagenesis in mouse hepatocytes, Genesis 26, 147–148.
68. Schuler, M., Dierich, A., Chambon, P., and Metzger, D. (2004) Efficient temporally controlled targeted somatic mutagenesis in hepatocytes of the mouse, Genesis 39, 167–172.
69. Kuhbandner, S., Brummer, S., Metzger, D., Chambon, P., Hofmann, F., and Feil, R. (2000) Temporally controlled somatic mutagenesis in smooth muscle, Genesis 28, 15–22.
70. Schuler, M., Ali, F., Metzger, E., Chambon, P., and Metzger, D. (2005) Temporally controlled targeted somatic mutagenesis in skeletal muscles of the mouse, Genesis 41, 165–170.
71. Sohal, D. S., Nghiem, M., Crackower, M. A., Witt, S. A., Kimball, T. R., Tymitz, K. M., Penninger, J. M., and Molkentin, J. D. (2001) Temporally regulated and tissue-specific gene manipulations in the adult and embryonic heart using a tamoxifen-inducible Cre protein, Circ Res 89, 20–25.
72. Kim, J. E., Nakashima, K., and de Crombrugghe, B. (2004) Transgenic mice expressing a ligand-inducible cre recombinase in osteoblasts and odontoblasts: a new tool to examine physiology and disease of postnatal bone and tooth, Am J Pathol 165, 1875–1882.
73. Chen, M., Lichtler, A. C., Sheu, T. J., Xie, C., Zhang, X., O’Keefe, R. J., and Chen, D. (2007) Generation of a transgenic mouse model with chondrocyte-specific and tamoxifen-inducible expression of Cre recombinase, Genesis 45, 44–50.
74. Imai, T., Jiang, M., Chambon, P., and Metzger, D. (2001) Impaired adipogenesis and lipolysis in the mouse upon selective ablation of the retinoid X receptor alpha mediated by a tamoxifen-inducible chimeric Cre recombinase (Cre-ERT2) in adipocytes, Proc Natl Acad Sci U S A 98, 224–228.
75. Hirrlinger, P. G., Scheller, A., Braun, C., Hirrlinger, J., and Kirchhoff, F. (2006) Temporal control of gene recombination in astrocytes by transgenic expression of the tamoxifen-inducible DNA recombinase variant CreERT2, Glia 54, 11–20.
76. Casper, K. B., Jones, K., and McCarthy, K. D. (2007) Characterization of astrocyte-specific conditional knockouts, Genesis 45, 292–299.
77. Mori, T., Tanaka, K., Buffo, A., Wurst, W., Kuhn, R., and Gotz, M. (2006) Inducible gene deletion in astroglia and radial glia – a valuable tool for functional and lineage analysis, Glia 54, 21–34.
78. Imayoshi, I., Ohtsuka, T., Metzger, D., Chambon, P., and Kageyama, R. (2006) Temporal regulation of Cre recombinase activity in neural stem cells, Genesis 44, 233–238.
79. Leone, D. P., Genoud, S., Atanasoski, S., Grausenburger, R., Berger, P., Metzger, D., Macklin, W. B., Chambon, P., and Suter, U. (2003) Tamoxifen-inducible glia-specific Cre mice for somatic mutagenesis in oligodendro-cytes and Schwann cells, Mol Cell Neurosci 22, 430–440.
80. Doerflinger, N. H., Macklin, W. B., and Popko, B. (2003) Inducible site-specific recombination in myelinating cells, Genesis 35, 63–72.
81. Gu, X., Yan, Y., Li, H., He, D., Pleasure, S. J., and Zhao, C. (2009) Characterization of the Frizzled10-CreER transgenic mouse: an inducible Cre line for the study of Cajal-Retzius cell development, Genesis 47, 210–216.
82. Zhao, J., Nassar, M. A., Gavazzi, I., and Wood, J. N. (2006) Tamoxifen-inducible NaV1.8-CreERT2 recombinase activity in nociceptive neurons of dorsal root ganglia, Genesis 44, 364–371.
83. el Marjou, F., Janssen, K. P., Chang, B. H., Li, M., Hindie, V., Chan, L., Louvard, D., Chambon, P., Metzger, D., and Robine, S. (2004) Tissue-specific and inducible Cre-mediated recombination in the gut epithelium, Genesis 39, 186–193.
84. Lantinga-van Leeuwen, I. S., Leonhard, W. N., van de Wal, A., Breuning, M. H., Verbeek, S., de Heer, E., and Peters, D. J. (2006) Transgenic mice expressing tamoxifen-inducible Cre for somatic gene modification in renal epithelial cells, Genesis 44, 225–232.
85. Wen, F., Cecena, G., Munoz-Ritchie, V., Fuchs, E., Chambon, P., and Oshima, R. G. (2003) Expression of conditional cre recombinase in epithelial tissues of transgenic mice, Genesis 35, 100–106.
86. Boyle, S., Misfeldt, A., Chandler, K. J., Deal, K. K., Southard-Smith, E. M., Mortlock, D. P., Baldwin, H. S., and de Caestecker, M. (2008) Fate mapping using Cited1-CreERT2 mice demonstrates that the cap mesenchyme contains self-renewing progenitor cells and gives rise exclusively to nephronic epithelia, Dev Biol 313, 234–245.
87. Forde, A., Constien, R., Grone, H. J., Hammerling, G., and Arnold, B. (2002) Temporal Cre-mediated recombination exclusively in endothelial cells using Tie2 regulatory elements, Genesis 33, 191–197.
88. Monvoisin, A., Alva, J. A., Hofmann, J. J., Zovein, A. C., Lane, T. F., and Iruela-Arispe, M. L. (2006) VE-cadherin-CreERT2 transgenic mouse: a model for inducible recombination in the endothelium, Dev Dyn 235, 3413–3422.
89. Claxton, S., Kostourou, V., Jadeja, S., Chambon, P., Hodivala-Dilke, K., and Fruttiger, M. (2008) Efficient, inducible Cre-recombinase activation in vascular endothelium, Genesis 46, 74–80.
90. Qiu, H. Y., Guo, C., Cheng, X. W., Huang, Y., Xiong, Z. Q., and Ding, Y. Q. (2008) Pitx3-CreER mice showing restricted Cre expression in developing ocular lens and skeletal muscles, Genesis 46, 324–328.