Analysis of legionella infection using RNAi in drosophila cells

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

Research output: ResearchChapter

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.

LanguageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages251-264
Number of pages14
Volume954
DOIs
StatePublished - 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume954
ISSN (Print)10643745

Fingerprint

Legionella
Double-Stranded RNA
RNA Interference
Drosophila
Infection
Growth
Legionnaires' Disease
Legionella pneumophila
RNA Stability
Gene Silencing
Culture Media
Cultured Cells
Cell Culture Techniques
Bacteria
Genes
Proteins

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

De Jesús, D. A., O'connor, T. J., & Isberg, R. R. (2013). Analysis of legionella infection using RNAi in drosophila cells. In Methods in Molecular Biology (Vol. 954, pp. 251-264). (Methods in Molecular Biology; Vol. 954). DOI: 10.1007/978-1-62703-161-5-15

Analysis of legionella infection using RNAi in drosophila cells. / De Jesús, Dennise A.; O'connor, Tamara J.; Isberg, Ralph R.

Methods in Molecular Biology. Vol. 954 2013. p. 251-264 (Methods in Molecular Biology; Vol. 954).

Research output: ResearchChapter

De Jesús, DA, O'connor, TJ & Isberg, RR 2013, Analysis of legionella infection using RNAi in drosophila cells. in Methods in Molecular Biology. vol. 954, Methods in Molecular Biology, vol. 954, pp. 251-264. DOI: 10.1007/978-1-62703-161-5-15
De Jesús DA, O'connor TJ, Isberg RR. Analysis of legionella infection using RNAi in drosophila cells. In Methods in Molecular Biology. Vol. 954. 2013. p. 251-264. (Methods in Molecular Biology). Available from, DOI: 10.1007/978-1-62703-161-5-15
De Jesús, Dennise A. ; O'connor, Tamara J. ; Isberg, Ralph R./ Analysis of legionella infection using RNAi in drosophila cells. Methods in Molecular Biology. Vol. 954 2013. pp. 251-264 (Methods in Molecular Biology).
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