Kalidas Savitha, Smith Dean P
Department of Pharmacology and Center for Basic Neuroscience, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA.
Neuron. 2002 Jan 17;33(2):177-84. doi: 10.1016/s0896-6273(02)00560-3.
Drosophila melanogaster has been a premier genetic model system for nearly 100 years, yet lacks a simple method to disrupt gene expression. Here, we show genomic cDNA fusions predicted to form double-stranded RNA (dsRNA) following splicing, effectively silencing expression of target genes in adult transgenic animals. We targeted three Drosophila genes: lush, white, and dGq(alpha). In each case, target gene expression is dramatically reduced, and the white RNAi phenotype is indistinguishable from a deletion mutant. This technique efficiently targets genes expressed in neurons, a tissue refractory to RNAi in C. elegans. These results demonstrate a simple strategy to knock out gene function in specific cells in living adult Drosophila that can be applied to define the biological function of hundreds of orphan genes and open reading frames.
近100年来,黑腹果蝇一直是首要的遗传模型系统,但缺乏一种简单的方法来干扰基因表达。在此,我们展示了基因组cDNA融合体,预测其在剪接后会形成双链RNA(dsRNA),能有效沉默成年转基因动物中靶基因的表达。我们针对三个果蝇基因:lush、white和dGq(α)。在每种情况下,靶基因表达都显著降低,且white RNAi表型与缺失突变体无法区分。该技术有效地靶向在神经元中表达的基因,而在秀丽隐杆线虫中,神经元组织对RNAi具有抗性。这些结果证明了一种在成年活体果蝇的特定细胞中敲除基因功能的简单策略,该策略可用于确定数百个孤儿基因和开放阅读框的生物学功能。