Abstract
Canonical RNAi, one of the so-called RNA-silencing mechanisms, is defined as sequence-specific RNA degradation induced by long double-stranded RNA (dsRNA). RNAi occurs in four basic steps: (i) processing of long dsRNA by RNase III Dicer into small interfering RNA (siRNA) duplexes, (ii) loading of one of the siRNA strands on an Argonaute protein possessing endonucleolytic activity, (iii) target recognition through siRNA basepairing, and (iv) cleavage of the target by the Argonaute’s endonucleolytic activity. This basic pathway diversified and blended with other RNA silencing pathways employing small RNAs. In some organisms, RNAi is extended by an amplification loop employing an RNA-dependent RNA polymerase, which generates secondary siRNAs from targets of primary siRNAs. Given the high specificity of RNAi and its presence in invertebrates, it offers an opportunity for highly selective pest control. The aim of this text is to provide an introductory overview of key mechanistic aspects of RNA interference for understanding its potential and constraints for its use in pest control.
Highlights
RNA interference (RNAi) is one of the pathways, collectively named RNA silencing pathways, that employ small RNAs as guides for sequence-specific silencing [reviewed in (Ketting, 2011)]
RNAi offers selective gene targeting in a species-specific manner
Off-targeting effects can be monitored in closely related species and targets and, if identified, they would disappear after termination of the RNAi treatment
Summary
RNA interference (RNAi) is one of the pathways, collectively named RNA silencing pathways, that employ small RNAs as guides for sequence-specific silencing [reviewed in (Ketting, 2011)]. For example, the basic RNAi mechanism has been integrated into a complex pathway system of post-transcriptional and transcriptional silencing, which employs multiple Dicer, Argonaute and RdRP proteins and functions in antiviral defense, protection of genome integrity, and regulation of gene expression [reviewed for example in (Bologna and Voinnet, 2014; Borges and Martienssen, 2015)]. Functional RNAi in mammalian cells requires high Dicer activity, enough dsRNA substrate, and suppression of the interferon response (Kennedy et al, 2015; Maillard et al, 2016; Kennedy et al, 2017; Van Der Veen et al, 2018; Demeter et al, 2019) These three conditions are rarely met—a unique example occurs in the mouse oocyte [reviewed in (Svoboda, 2014)]. In one of the plant RNA silencing mechanisms, RNAi essentially serves as an amplifier of miRNA silencing where miRNA-mediated cleavage of mRNA targets is followed by RdRP-mediated production of long dsRNA, which is processed by Dicer into so-called phased siRNAs (phasiRNA)
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