Abstract

Double-stranded RNA (dsRNA) molecules of viral origin trigger a post-transcriptional gene-silencing mechanism called RNA interference (RNAi). Specifically, virally derived dsRNA is recognized and cleaved by the enzyme Dicer2 into short interfering RNAs (siRNAs), which further direct sequence-specific RNA silencing, ultimately silencing replication of the virus. Notably, RNAi can also be artificially triggered by the delivery of gene-specific dsRNA, thereby leading to endogenous gene silencing. This is a widely used technology that holds great potential to contribute to novel pest control strategies. In this regard, research efforts have been set to find methods to efficiently trigger RNAi in the field. In this article, we demonstrate the generation of dsRNA- and/or virus-derived siRNAs—the main RNAi effectors—in six insect species belonging to five economically important orders (Lepidoptera, Orthoptera, Hymenoptera, Coleoptera, and Diptera). In addition, we describe that the siRNA length distribution is species-dependent. Taken together, our results reveal interspecies variability in the (antiviral) RNAi mechanism in insects and show promise to contribute to future research on (viral-based) RNAi-triggering mechanisms in this class of animals.

Highlights

  • RNA interference (RNAi) is a post-transcriptional gene-silencing mechanism triggered by double-stranded RNA molecules

  • We describe the species-dependent length distribution of Double-stranded RNA (dsRNA)- and/or virus-derived short interfering RNAs (siRNAs) in six insect species belonging to five orders: Spodoptera exigua and Trichoplusia ni (Lepidoptera); Locusta migratoria (Orthoptera); Bombus terrestris (Hymenoptera); Tribolium castaneum (Coleoptera); and Drosophila melanogaster (Diptera)

  • In order to investigate the generation of Cricket Paralysis Virus (CrPV)-specific siRNAs, a CrPV infection was induced in five distinct insect species belonging to economically relevant orders

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Summary

Introduction

RNA interference (RNAi) is a post-transcriptional gene-silencing mechanism triggered by double-stranded RNA (dsRNA) molecules. As these RNA duplexes are mainly produced during the replicative cycle of viruses, RNAi is an important antiviral response. In insects, this pathway can be described in three central steps. The trigger dsRNA is recognized by the RNase III enzyme. Dicer and processed into small RNA duplexes called small interfering RNAs (siRNAs). These siRNAs (18–24 nt long) are loaded into the RNA-induced silencing complex (RISC) and unwound.

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