Rapid construction of Drosophila RNAi transgenes using pRISE, a P-element-mediated transformation vector exploiting an in vitro recombination system

• Published in: Genes & Genetic Systems Vol. 81; no. 2; pp. 129 - 134
• Main Authors: Kondo, Takefumi, Inagaki, Sachi, Yasuda, Kunio, Kageyama, Yuji
• Format: Journal Article
• Language: English
• Published: Japan The Genetics Society of Japan 2006; Japan Science and Technology Agency
• Subjects: Animals; Animals, Genetically Modified; ATP-Binding Cassette Transporters - metabolism; Cells, Cultured; DNA Transposable Elements - genetics; Drosophila; Drosophila - genetics; Drosophila Proteins - deficiency; Drosophila Proteins - metabolism; Eye Proteins - metabolism; Gateway Technology; Genetic Vectors - chemical synthesis; Recombination, Genetic; RNA Interference; RNA, Small Interfering - genetics; RNAi; transformation vector; Transformation, Genetic; Transgenes
• ISSN: 1341-7568; 1880-5779
• DOI: 10.1266/ggs.81.129

RNAi is a gene-silencing phenomenon mediated by double-stranded RNA (dsRNA) and has become a powerful tool to elucidate gene function. To accomplish rapid construction of transgenes expressing dsRNA in Drosophila, we developed a novel transformation vector, pRISE, which contains an inverted repeat of the attR1-ccdB-attR2 cassette for in vitro recombination and a pentameric GAL4 binding site for conditional expression. These features enabled us to construct RNAi transgenes without a complicated cloning scheme. In cultured cells and transgenic flies, pRISE constructs carrying dsRNA transgenes induced effective RNAi against an EGFP transgene and the endogenous white gene, respectively. These results indicate that pRISE is a convenient transformation vector for studies of multiple Drosophila genes for which functional information is lacking.