基于 dsRNA 摄食的褐飞虱（半翅目：飞虱科）中的 RNA 干扰。
RNA interference in Nilaparvata lugens (Homoptera: Delphacidae) based on dsRNA ingestion.
机构信息
Hubei Insect Resources Utilisation and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.
出版信息
Pest Manag Sci. 2011 Jul;67(7):852-9. doi: 10.1002/ps.2124. Epub 2011 Mar 2.
BACKGROUND
An efficient and convenient RNA interference (RNAi) technique involving double-stranded RNA (dsRNA) ingestion is useful for gene function studies of non-model insects.
RESULTS
Three dsRNAs targeting different sites within a gene encoding vacuolar ATP synthase subunit E (V-ATPase-E, 21E01) were synthesised for RNAi in Nilaparvata lugens. dsRNA was found to be stable in 0.1 g mL(-1) sucrose solution, but unstable in artificial fodder. Therefore, dsRNAs were orally delivered into N. lugens in 0.1 g mL(-1) sucrose solution. RNAi was induced by all three of the dsRNAs at 0.05 µg µL(-1) in N. lugens. Time dynamics analysis of gene silencing indicated that significant suppression of the target gene began as early as 2 days after ingestion of ds2-21E01 and ds3-21E01. However, significant repressive effects were recorded up to 10 days after exposure to ds1-21E01. The maximum reduction in target gene mRNA was observed after 10 days of treatment, with suppression ratios induced by ds1-21E01, ds2-21E01 and ds3-21E01 of 41, 55 and 48% respectively.
CONCLUSION
An efficient and convenient RNAi technique involving dsRNA ingestion has been successfully developed for N. lugens. This will be a useful tool for further functional genomic investigation in this organism.
背景
涉及双链 RNA（dsRNA）摄入的高效便捷 RNA 干扰（RNAi）技术，对于非模式昆虫的基因功能研究非常有用。
结果
针对编码液泡型 ATP 合酶亚基 E（V-ATPase-E，21E01）的基因内不同位点，合成了 3 种 dsRNA 用于褐飞虱的 RNAi。dsRNA 在 0.1 g mL（-1）蔗糖溶液中稳定，但在人工饲料中不稳定。因此，dsRNA 以 0.1 g mL（-1）蔗糖溶液的形式口服递送至褐飞虱体内。用三种 dsRNA 中的任意一种，浓度为 0.05 µg µL（-1）时，都可以在褐飞虱中诱导 RNAi。基因沉默的时间动态分析表明，在摄入 ds2-21E01 和 ds3-21E01 后 2 天，靶基因就开始出现显著抑制。然而，在接触 ds1-21E01 后 10 天内仍记录到显著的抑制效果。处理 10 天后，靶基因 mRNA 最大程度减少，ds1-21E01、ds2-21E01 和 ds3-21E01 诱导的抑制率分别为 41%、55%和 48%。
结论
成功开发了一种涉及 dsRNA 摄入的高效便捷 RNAi 技术，用于褐飞虱。这将成为该生物进一步进行功能基因组研究的有用工具。
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