RNA interference (RNAi) can be used to silence genes in a number of species, including plants, nematodes, protozoans, Drosophila melanogaster, mouse embryos, and mammalian and Drosophila cell cultures. Drosophila cell culture provides the opportunity to study signal transduction pathways and protein function in a simple, well defined cell culture paradigm. Furthermore, because Drosophila are RNAi responsive, the results obtained from experiments performed on cultured cells can be confirmed in the whole organism. RNAi takes advantage of the unique ability of double-stranded RNA (dsRNA) molecules to induce gene silencing in a highly specific manner. This phenomenon is efficacious and long lived, being passed to subsequent generations in Drosophila cell culture. To date, many Drosophila cell lines tested respond to dsRNAs by ablating expression of the target protein. Furthermore, all dsRNAs tested have been efficacious at silencing the target gene. Drosophila cell cultures are simple, easily manipulated model systems that will facilitate loss of function studies applicable to a wide variety of questions.