RNAi effects on actin mRNAs in Homalodisca vitripennis cells.

Cristina Rosa, Shizuo G Kamita, Haley Dequine, Ha Wuriyanghan, John A Lindbo, Bryce W Falk
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Abstract

The xylem feeding leafhopper Homalodisaca vitripennis (H. vitripennis) is an unusually robust and efficient vector of Xylella fastidiosa, a Gram-negative bacterium which causes several very important plant diseases. Here we investigated RNA interference (RNAi) to target actin, a key component of insect cells and whole bodies, in H. vitripennis cells. RNAi effectors were delivered via lipid based transfection and real-time RT-PCR, RNA hybridization, and microscopic analyses were employed to verify RNAi effects. When actin dsRNAs were used, a 10-fold decrease in the target H. vitripennis actin mRNA level was seen in cells. Altered phenotypic effects also were evident in transfected cells, as were small interfering RNAs, hallmarks of RNAi. The use of H. vitripennis cells and RNAi offers new opportunities to research hemipterans, the most important insect vectors of plant pathogens.

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RNAi 对同种异形虫细胞肌动蛋白 mRNA 的影响
木质部食叶蝉(Homalodisaca vitripennis,H. vitripennis)是 Xylella fastidiosa(一种革兰氏阴性细菌,可导致多种非常重要的植物病害)的一种异常强大而高效的载体。在这里,我们研究了以肌动蛋白(昆虫细胞和整体的关键组成部分)为靶标的 RNA 干扰(RNAi)。我们通过脂质转染传递 RNAi 效应子,并采用实时 RT-PCR、RNA 杂交和显微镜分析来验证 RNAi 的效果。当使用肌动蛋白 dsRNA 时,细胞中的目标 H. vitripennis 肌动蛋白 mRNA 水平下降了 10 倍。转染细胞的表型效应也发生了明显的改变,小干扰 RNA 也是 RNAi 的标志。利用H. vitripennis细胞和RNAi为研究植物病原体最重要的昆虫载体--半翅目昆虫提供了新的机会。
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