Thairu M W, Skidmore I H, Bansal R, Nováková E, Hansen T E, Li-Byarlay H, Wickline S A, Hansen A K
Department of Entomology, University of Illinois, Urbana-Champaign, IL, USA.
Department of Entomology, The Ohio State University, Wooster, OH, USA.
Insect Mol Biol. 2017 Jun;26(3):356-368. doi: 10.1111/imb.12301. Epub 2017 Mar 17.
RNA interference (RNAi) has emerged as a promising method for validating gene function; however, its utility in nonmodel insects has proven problematic, with delivery methods being one of the main obstacles. This study investigates a novel method of RNAi delivery in aphids, the aerosolization of short interfering RNA (siRNA)-nanoparticle complexes. By using nanoparticles as a siRNA carrier, the likelihood of cellular uptake is increased, when compared to methods previously used in insects. To determine the efficacy of this RNAi delivery system, siRNAs were aerosolized with and without nanoparticles in three aphid species: Acyrthosiphon pisum, Aphis glycines and Schizaphis graminum. The genes targeted for knockdown were carotene dehydrogenase (tor), which is important for pigmentation in Ac. pisum, and branched chain-amino acid transaminase (bcat), which is essential in the metabolism of branched-chain amino acids in all three aphid species. Overall, we observed modest gene knockdown of tor in Ac. pisum and moderate gene knockdown of bcat in Ap. glycines along with its associated phenotype. We also determined that the nanoparticle emulsion significantly increased the efficacy of gene knockdown. Overall, these results suggest that the aerosolized siRNA-nanoparticle delivery method is a promising new high-throughput and non-invasive RNAi delivery method in some aphid species.
RNA干扰（RNAi）已成为一种验证基因功能的有前景的方法；然而，事实证明它在非模式昆虫中的应用存在问题，其中递送方法是主要障碍之一。本研究调查了一种在蚜虫中递送RNAi的新方法，即短干扰RNA（siRNA）-纳米颗粒复合物的气溶胶化。与先前在昆虫中使用的方法相比，通过使用纳米颗粒作为siRNA载体，细胞摄取的可能性增加。为了确定这种RNAi递送系统的功效，在三种蚜虫物种（豌豆蚜、大豆蚜和麦二叉蚜）中，分别在有纳米颗粒和无纳米颗粒的情况下对siRNA进行气溶胶化处理。被靶向敲低的基因是胡萝卜素脱氢酶（tor），它对豌豆蚜的色素沉着很重要，以及支链氨基酸转氨酶（bcat），它在所有三种蚜虫物种的支链氨基酸代谢中都必不可少。总体而言，我们观察到豌豆蚜中tor基因有适度的敲低，大豆蚜中bcat基因有中度的敲低及其相关表型。我们还确定纳米颗粒乳液显著提高了基因敲低的功效。总体而言，这些结果表明，气溶胶化的siRNA-纳米颗粒递送方法是一种在某些蚜虫物种中很有前景的新型高通量且非侵入性的RNAi递送方法。
