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

被引：20

作者：

Kondo, Takefumi ^{[1
]}

Inagaki, Sachi ^{[1
]}

Yasuda, Kunio ^{[1
]}

Kageyama, Yuji ^{[1
]}

机构：

[1] Nara Inst Sci & Technol, Grad Sch Biol Sci, Ikoma 6300192, Japan

来源：

GENES & GENETIC SYSTEMS | 2006年 / 81卷 / 02期

关键词：

Drosophila; gateway technology; RNAi; transformation vector;

D O I：

10.1266/ggs.81.129

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：129 / 134

页数：6

共 27 条

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