Summary
Innate immunity is an ancient and conserved defense mechanism. The worm Caenorhabditis elegans provides a useful tool for studying the function of the innate immune system at the molecular and cellular levels within the context of a whole organism. The powerful genetics of the worm, combined with efficacy of gene knockdown by RNA interference (RNAi), offer complementary tools for analyzing the contribution of individual genes to innate immunity. It is important, however, to exclude pleiotropic effects that confound results. In this chapter, we will describe the procedures for performing both forward and reverse genetic screens and will discuss a number of techniques developed to resolve confounding effects, thus enhancing the power of this system.
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
Hoffmann, J. A., Kafatos, F. C., Janeway, C. A., and Ezekowitz, R. A. (1999) Phylogenetic perspectives in innate immunity. Science 284, 1313–8.
Janeway, C. A., Jr., and Medzhitov, R. (2002) Innate immune recognition. Annu Rev Immunol 20, 197–216.
Tan, M. W., Mahajan-Miklos, S., and Ausubel, F. M. (1999) Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis. Proc Natl Acad Sci USA 96, 715–20.
Alegado, R. A., Campbell, M. C., Chen, W. C., Slutz, S. S., and Tan, M. W. (2003) Characterization of mediators of microbial virulence and innate immunity using the Caenorhabditis elegans host-pathogen model. Cell Microbiol 5, 435–44.
Tan, M. W., Rahme, L. G., Sternberg, J. A., Tompkins, R. G., and Ausubel, F. M. (1999) Pseudomonas aeruginosa killing of Caenorhabditis elegans used to identify P. aeruginosa virulence factors. Proc Natl Acad Sci USA 96, 2408–13.
Kim, D. H., Feinbaum, R., Alloing, G., Emerson, F. E., Garsin, D. A., Inoue, H., Tanaka-Hino, M., Hisamoto, N., Matsumoto, K., Tan, M. W., and Ausubel, F. M. (2002) A conserved p38 MAP kinase pathway in Caenorhabditis elegans innate immunity. Science 297, 623–6.
Timmons, L., and Fire, A. (1998) Specific interference by ingested dsRNA. Nature 395, 854.
Guo, S., and Kemphues, K. J. (1995) par-1, a gene required for establishing polarity in C. elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed. Cell 81, 611–20.
Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., and Mello, C. C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–11.
Kamath, R. S., Fraser, A. G., Dong, Y., Poulin, G., Durbin, R., Gotta, M., Kanapin, A., Le Bot, N., Moreno, S., Sohrmann, M., Welchman, D. P., Zipperlen P., and Ahringer, J. (2003) Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 421, 231–7.
Grishok, A. (2005) RNAi mechanisms in Caenorhabditis elegans. FEBS Lett 579, 5932–9.
Rahme, L. G., Stevens, E. J., Wolfort, S. F., Shao, J., Tompkins, R. G., and Ausubel, F. M. (1995) Common virulence factors for bacterial pathogenicity in plants and animals. Science 268, 1899–902.
Labrousse, A., Chauvet, S., Couillault, C., Kurz, C. L., and Ewbank, J. J. (2000) Caenorhabditis elegans is a model host for Salmonella typhimurium. Curr Biol 10, 1543–5.
Brenner, S. (1974) The genetics of Caenorhabditis elegans. Genetics 77, 71–94.
Gems, D., and Riddle, D. L. (2000) Genetic, behavioral and environmental determinants of male longevity in Caenorhabditis elegans. Genetics 154, 1597–610.
Thomas, J. H. (1990) Genetic analysis of defecation in Caenorhabditis elegans. Genetics 124, 855–72.
Liu, D. W., and Thomas, J. H. (1994) Regulation of a periodic motor program in C. elegans. J Neurosci 14, 1953–62.
Iwasaki, K., Liu, D. W., and Thomas, J. H. (1995) Genes that control a temperature-compensated ultradian clock in Caenorhabditis elegans. Proc Natl Acad Sci USA 92, 10317–21.
Hodgkin, J. A., and Brenner, S. (1977) Mutations causing transformation of sexual phenotype in the nematode Caenorhabditis elegans. Genetics 86, 275–87.
Beanan, M. J., and Strome, S. (1992) Characterization of a germ-line proliferation mutation in C. elegans. Development 116, 755–66.
Simmer, F., Tijsterman, M., Parrish, S., Koushika, S. P., Nonet, M. L., Fire, A., Ahringer, J., and Plasterk, R. H. (2002) Loss of the putative RNA-directed RNA polymerase RRF-3 makes C. elegans hypersensitive to RNAi. Curr Biol 12, 1317–9.
Ashcroft, N. R., Srayko, M., Kosinski, M. E., Mains, P. E., and Golden, A. (1999) RNA-Mediated interference of a cdc25 homolog in Caenorhabditis elegans results in defects in the embryonic cortical membrane, meiosis, and mitosis. Dev Biol 206, 15–32.
Ashcroft, N., and Golden, A. (2002) CDC-25.1 regulates germline proliferation in Caenorhabditis elegans. Genesis 33, 1–7.
Varkey, J. P., Muhlrad, P. J., Minniti, A. N., Do, B., and Ward, S. (1995) The Caenorhabditis elegans spe-26 gene is necessary to form spermatids and encodes a protein similar to the actin-associated proteins kelch and scruin. Genes Dev 9, 1074–86.
Reiner, D. J., and Thomas, J. H. (1995) Reversal of a muscle response to GABA during C. elegans male development. J Neurosci 15, 6094–102.
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
Shapira, M., Tan, MW. (2008). Genetic Analysis of Caenorhabditis elegans Innate Immunity. 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_25
Download citation
DOI: https://doi.org/10.1007/978-1-59745-570-1_25
Publisher Name: Humana Press
Print ISBN: 978-1-58829-746-4
Online ISBN: 978-1-59745-570-1
eBook Packages: Springer Protocols