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Investigating the Involvement of Host Factors Involved in Intracellular Pathogen Infection by RNAi in Drosophila Cells

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Innate Immunity

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 415))

Summary

Intracellular pathogens represent a serious threat to human health. Although the biology and the virulence factors involved in intracellular bacterial infection are relatively well documented, little is known about the host factors involved in the infection process. This situation is mainly due to the difficulty of conducting extensive genetic analyses in the targeted host cells and points to the need for developing genetic systems to model the infection process. Here, we describe a method (amenable to high-throughput analysis) that allows for the study of host factors involved in intracellular pathogen infection by using dsRNA-mediated gene expression knockdown [RNA interference (RNAi)] in Drosophila cell lines.

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References

  1. Samakovlis, C., Asling, B., Boman, H. G., Gateff, E., and Hultmark, D. (1992) In vitro induction of cecropin genes–an immune response in a Drosophila blood cell line. Biochem Biophys Res Commun 188, 1169–75

    Article  CAS  PubMed  Google Scholar 

  2. Boutros, M., Agaisse, H., and Perrimon, N. (2002) Sequential activation of signaling pathways during innate immune responses in Drosophila. Dev Cell 3, 711–22.

    Article  CAS  PubMed  Google Scholar 

  3. Silverman, N., Zhou, R., Erlich, R. L., Hunter, M., Bernstein, E., Schneider, D., and Maniatis, T. (2003) Immune activation of NF-kappaB and JNK requires Drosophila TAK1. J Biol Chem 278, 48928–34.

    Article  CAS  PubMed  Google Scholar 

  4. Cheng, L. W., Viala, J. P., Stuurman, N., Wiedemann, U., Vale, R. D., and Portnoy, D. A. (2005) Use of RNA interference in Drosophila S2 cells to identify host pathways controlling compartmentalization of an intracellular pathogen. Proc Natl Acad Sci USA 102, 13646–51.

    Article  CAS  PubMed  Google Scholar 

  5. Mansfield, B. E., Dionne, M. S., Schneider, D. S., and Freitag, N. E. (2003) Exploration of host-pathogen interactions using Listeria monocytogenes and Drosophila melanogaster. Cell Microbiol 5, 901–11.

    Article  CAS  PubMed  Google Scholar 

  6. Clemens, J. C., Worby, C. A., Simonson-Leff, N., Muda, M., Maehama, T., Hemmings, B. A., and Dixon, J. E. (2000) Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways. Proc Natl Acad Sci USA 97, 6499–503.

    Article  CAS  PubMed  Google Scholar 

  7. Agaisse, H., Burrack, L. S., Philips, J. A., Rubin, E. J., Perrimon, N., and Higgins, D. E. (2005) Genome-wide RNAi screen for host factors required for intracellular bacterial infection. Science 309, 1248–51.

    Article  CAS  PubMed  Google Scholar 

  8. 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.

    Article  CAS  PubMed  Google Scholar 

  9. Portnoy, D. A., Auerbuch, V., and Glomski, I. J. (2002) The cell biology of Listeria monocytogenes infection: the intersection of bacterial pathogenesis and cell-mediated immunity. J Cell Biol 158, 409–14.

    Article  CAS  PubMed  Google Scholar 

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© 2008 Humana Press Inc.

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Agaisse, H. (2008). Investigating the Involvement of Host Factors Involved in Intracellular Pathogen Infection by RNAi in Drosophila Cells. 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_23

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  • DOI: https://doi.org/10.1007/978-1-59745-570-1_23

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-746-4

  • Online ISBN: 978-1-59745-570-1

  • eBook Packages: Springer Protocols

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