Institute of Life Science, Swansea University Medical School, Singleton Park, Swansea, UK.
Methods Mol Biol. 2022;2360:295-306. doi: 10.1007/978-1-0716-1633-8_21.
RNA interference (RNAi) has emerged as a widely used approach for reverse genetic analysis in eukaryotes. In insects, RNAi also has an application in the control of insect pests. Several methods have been developed for delivery of interfering RNA in insects, with varying outcomes for different species. Here we describe how a bacterial symbiont can be exploited for continuous synthesis of interfering double-stranded RNA (dsRNA) in its insect host. This approach, termed symbiont-mediated RNAi (SMR), can overcome problems associated with instability of dietary dsRNA due to action of salivary or foregut nucleases. As insects do not possess RNA-dependent RNA polymerase activity that can amplify and extend RNAi in other organisms, SMR also offers the possibility of long-term systemic RNAi not afforded by single applications of dsRNA to insects by other delivery methods. Here, we describe how SMR can be applied in a globally distributed agricultural pest species, western flower thrips (Frankliniella occidentalis).
RNA 干扰 (RNAi) 已成为真核生物反向遗传学分析的一种广泛应用的方法。在昆虫中，RNAi 也可用于防治害虫。已经开发出几种在昆虫中递送干扰 RNA 的方法，对于不同的物种有不同的效果。在这里，我们描述了如何利用一种细菌共生体在其昆虫宿主中连续合成干扰双链 RNA (dsRNA)。这种方法称为共生介导的 RNAi (SMR)，可以克服由于唾液或前肠核酸酶的作用导致膳食 dsRNA 不稳定的问题。由于昆虫不具有能够在其他生物体中扩增和扩展 RNAi 的 RNA 依赖性 RNA 聚合酶活性，因此 SMR 还提供了通过其他递送方法单次应用 dsRNA 无法实现的长期系统性 RNAi 的可能性。在这里，我们描述了如何将 SMR 应用于一种分布广泛的农业害虫物种，即西部花蓟马（Frankliniella occidentalis）。
