Rapid construction of Drosophila RNAi transgenes using pRISE, a P-element-mediated transformation vector exploiting an in vitro recombination system
Rapid construction of Drosophila RNAi transgenes using pRISE, a P-element-mediated transformation vector exploiting an in vitro recombination system
复制标题
DOI：
10.1266/ggs.81.129
复制
发表时间：
2006-04-01
影响因子：
1.1
通讯作者：
Kageyama, Yuji
中科院分区：
文献类型：
--
作者：
Kondo, Takefumi;Inagaki, Sachi;Kageyama, Yuji
RNAi是一种由双链RNA（dsRNA）介导的基因沉默现象，已成为研究基因功能的有力工具。为了实现在果蝇中快速构建表达dsRNA的转基因，我们开发了一种新的转化载体，pRISE，其包含用于体外重组的attR 1-ccdB-attR 2盒的反向重复序列和用于条件表达的五聚体GAL 4结合位点。这些特征使我们能够构建RNAi转基因，而无需复杂的克隆方案。在培养的细胞和转基因果蝇中，携带dsRNA转基因的pRISE构建体分别诱导针对EGFP转基因和内源性白色基因的有效RNAi。这些结果表明，pRISE是一个方便的转化载体，用于研究多个果蝇基因的功能信息缺乏。
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.