Analysis of legionella infection using RNAi in drosophila cells

Dennise A. De Jesús, Tamara J. O'connor, Ralph R. Isberg

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

RNA interference (RNAi) is the process of specific gene silencing by the use of double-stranded RNA (dsRNA). In cultured Drosophila cells, RNAi methodologies are well established and easily executed: dsRNA, when added to the cell culture medium, is efficiently internalized by the cells and, through the activity of endogenous processing machinery, targets the specified mRNA for degradation resulting in reduced levels of its encoded protein. This technique has proven very useful in studying the role of host genes during Legionella pneumophila infections, as it allows the effect of host factor depletion on intracellular growth of the bacterium to be examined. In this chapter we present the methods commonly used in our laboratory to study intracellular growth of L. pneumophila using dsRNA in Drosophila cells.

Original languageEnglish (US)
Title of host publicationLegionella
Subtitle of host publicationMethods and Protocols
EditorsCarmen Buchrieser, Hubert Hilbi
Pages251-264
Number of pages14
DOIs
StatePublished - Jan 14 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume954
ISSN (Print)1064-3745

Keywords

  • Drosophila melanogaster
  • Kc167
  • Legionella
  • RNA interference
  • dsRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    De Jesús, D. A., O'connor, T. J., & Isberg, R. R. (2013). Analysis of legionella infection using RNAi in drosophila cells. In C. Buchrieser, & H. Hilbi (Eds.), Legionella: Methods and Protocols (pp. 251-264). (Methods in Molecular Biology; Vol. 954). https://doi.org/10.1007/978-1-62703-161-5-15