title: Analyzing the Response of RNAi-Treated Drosophila Cells to Death Stimuli by Quantitative Real-Time Polymerase Chain Reaction.
abstract: A useful complement to animal studies is the use of Drosophila cell lines to analyze cell-death responses. There are numerous Drosophila cell lines available, such as S2 cells, which possess the advantages of being semi-adherent, fast growing, relatively robust, and useful for transfection and knockdown studies, whereas other lines, such as mbn2, are more suitable for analyzing hormone-induced cell death and gene expression. Drosophila cell lines are very amenable to knockdown studies as the cells take up double-stranded RNA (dsRNA) from the medium, initiating gene silencing and resulting in a high level of gene knockdown. This means that the cell lines are useful for investigating the response to death stimuli, following gene knockdown, by examining the expression of cell-death genes. This protocol describes the synthesis of dsRNA for treatment of Drosophila cells and the subsequent analysis of cell-death gene expression by quantitative real-time polymerase chain reaction (qPCR).
authors.affiliation: Centre for Cancer Biology, University of South Australia, Adelaide, South Australia 5001, Australia.
authors.forename: Donna
authors.initials: D
authors.lastname: Denton
authors.affiliation: Centre for Cancer Biology, University of South Australia, Adelaide, South Australia 5001, Australia.
authors.forename: Sharad
authors.initials: S
authors.lastname: Kumar
mesh_terms: D000818:Animals; D016923:Cell Death / Q000187:drug effects*; D002460:Cell Line; D004330:Drosophila; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*