Wynant N, Santos D, Van Wielendaele P, Vanden Broeck J
Molecular Developmental Physiology and Signal Transduction, Department of Animal Physiology and Neurobiology, KU Leuven, Leuven, Belgium.
Insect Mol Biol. 2014 Jun;23(3):320-9. doi: 10.1111/imb.12083. Epub 2014 Feb 14.
RNA interference (RNAi) has become a widely used loss-of-function tool in eukaryotes; however, the delivery of double-stranded (ds)RNA) to the target cells remains a major challenge when exploiting the RNAi-technology. In insects, the efficiency of RNAi is highly species-dependent. Yet, the mechanism of cell entry in insects has only been characterized in a cell line of the fruit fly, Drosophila melanogaster, a species that is well known to be poorly amenable to environmental RNAi. In the present paper, we demonstrate that silencing vacuolar H-ATPase 16 (vha16) and clathrin heavy chain (clath), two components of the Clathrin-dependent endocytosis pathway, together with pharmacological inhibition of scavenger receptors with polyinosine and dextran sulphate, can significantly attenuate the highly robust RNAi response in the desert locust, Schistocerca gregaria.
RNA干扰(RNAi)已成为真核生物中广泛使用的功能丧失工具;然而,在利用RNAi技术时,将双链(ds)RNA递送至靶细胞仍然是一个主要挑战。在昆虫中,RNAi的效率高度依赖物种。然而,昆虫细胞进入机制仅在果蝇(Drosophila melanogaster)的一个细胞系中得到表征,果蝇是一种众所周知的对环境RNAi不太敏感的物种。在本文中,我们证明,沉默网格蛋白依赖性内吞途径的两个组分——液泡H-ATP酶16(vha16)和网格蛋白重链(clath),以及用聚肌苷和硫酸葡聚糖对清道夫受体进行药理学抑制,可显著减弱沙漠蝗(Schistocerca gregaria)中高度强烈的RNAi反应。