Summary
Forward genetics has led to many “breakthrough” discoveries, and with the mouse genome almost fully sequenced, the creation of phenotypes through random germline mutagenesis has become an efficient means by which to find the function of yet undescribed genes. In this chapter, we will provide a practical guideline for performing germline mutagenesis in mice. In particular, we will focus on the application of this technology to identify genes that are essential to innate immune defense.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Beck, J. A., Lloyd, S., Hafezparast, M., Lennon-Pierce, M., Eppig, J. T., Festing, M. F., & Fisher, E. M. (2000). Genealogies of mouse inbred strains. Nat. Genet. 24, 23–25.
Guenet, J. L. & Bonhomme, F. (2003). Wild mice: an ever-increasing contribution to a popular mammalian model. Trends Genet. 19, 24–31.
Brunkow, M. E., Jeffery, E. W., Hjerrild, K. A., Paeper, B., Clark, L. B., Yasayko, S. A., Wilkinson, J. E., Galas, D., Ziegler, S. F., & Ramsdell, F. (2001). Disruption of a new forkhead/winged-helix protein, scurfin, results in the fatal lymphoproliferative disorder of the scurfy mouse. Nat. Genet. 27, 68–73.
Lee, S. H., Webb, J. R., & Vidal, S. M. (2002). Innate immunity to cytomegalovirus: the Cmv1 locus and its role in natural killer cell function. Microbes. Infect. 4, 1491–1503.
Poltorak, A., He, X., Smirnova, I., Liu, M. Y., Van Huffel, C., Du, X., Birdwell, D., Alejos, E., Silva, M., Galanos, C., Freudenberg, M., Ricciardi-Castagnoli, P., Layton, B., & Beutler, B. (1998). Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282, 2085–2088.
Scalzo, A. A., Wheat, R., Dubbelde, C., Stone, L., Clark, P., Du, Y., Dong, N., Stoll, J., Yokoyama, W. M., & Brown, M. G. (2003). Molecular genetic characterization of the distal NKC recombination hotspot and putative murine CMV resistance control locus. Immunogenetics 55, 370–378.
Webb, J. R., Lee, S. H., & Vidal, S. M. (2002). Genetic control of innate immune responses against cytomegalovirus: MCMV meets its match. Genes Immun. 3, 250–262.
Balling, R. (2001). ENU mutagenesis: analyzing gene function in mice. Annu. Rev. Genomics Hum. Genet. 2, 463–492.
Nusslein-Volhard, C. & Wieschaus, E. (1980). Mutations affecting segment number and polarity in Drosophila. Nature 287, 795–801.
Favor, J., Neuhauser-Klaus, A., & Ehling, U. H. (1988). The effect of dose fractionation on the frequency of ethylnitrosourea-induced dominant cataract and recessive specific locus mutations in germ cells of the mouse. Mutat. Res. 198, 269–275.
Favor, J., Neuhauser-Klaus, A., Ehling, U. H., Wulff, A., & van Zeeland, A. A. (1997). The effect of the interval between dose applications on the observed specific-locus mutation rate in the mouse following fractionated treatments of spermatogonia with ethylnitrosourea. Mutat. Res. 374, 193–199.
Rinchik, E. M., Carpenter, D. A., & Selby, P. B. (1990). A strategy for fine-structure functional analysis of a 6- to 11-centimorgan region of mouse chromosome 7 by high-efficiency mutagenesis. Proc Natl. Acad. Sci. U. S. A 87, 896–900.
Rinchik, E. M. & Carpenter, D. A. (1999). N-ethyl-N-nitrosourea mutagenesis of a 6- to 11-cM subregion of the Fah-Hbb interval of mouse chromosome 7: completed testing of 4557 gametes and deletion mapping and complementation analysis of 31 mutations. Genetics 152, 373–383.
Russell, W. L., Hunsicker, P. R., Raymer, G. D., Steele, M. H., Stelzner, K. F., & Thompson, H. M. (1982). Dose–response curve for ethylnitrosourea-induced specific-locus mutations in mouse spermatogonia. Proc Natl. Acad. Sci. U. S. A 79, 3589–3591.
Russell, W. L., Hunsicker, P. R., Carpenter, D. A., Cornett, C. V., & Guinn, G. M. (1982). Effect of dose fractionation on the ethylnitrosourea induction of specific-locus mutations in mouse spermatogonia. Proc Natl. Acad. Sci. U. S. A 79, 3592–3593.
Justice, M. J., Noveroske, J. K., Weber, J. S., Zheng, B., & Bradley, A. (1999). Mouse ENU mutagenesis. Hum. Mol. Genet. 8, 1955–1963.
Concepcion, D., Seburn, K. L., Wen, G., Frankel, W. N., & Hamilton, B. A. (2004). Mutation rate and predicted phenotypic target sizes in ethylnitrosourea-treated mice. Genetics 168, 953–959.
Hoebe, K., Du, X., Goode, J., Mann, N., & Beutler, B. (2003). Lps2: a new locus required for responses to lipopolysaccharide, revealed by germline mutagenesis and phenotypic screening. J. Endotoxin. Res. 9, 250–255.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press Inc.
About this protocol
Cite this protocol
Georgel, P., Du, X., Hoebe, K., Beutler, B. (2008). ENU Mutagenesis in Mice. In: Ewbank, J., Vivier, E. (eds) Innate Immunity. Methods in Molecular Biology™, vol 415. Humana Press. https://doi.org/10.1007/978-1-59745-570-1_1
Download citation
DOI: https://doi.org/10.1007/978-1-59745-570-1_1
Publisher Name: Humana Press
Print ISBN: 978-1-58829-746-4
Online ISBN: 978-1-59745-570-1
eBook Packages: Springer Protocols