Abstract
Artificial small RNAs (art-sRNAs), such as artificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs), are highly specific 21-nucleotide small RNAs designed to recognize and silence complementary target RNAs. Art-sRNAs are extensively used in gene function studies or for improving crops, particularly to protect plants against viruses. Typically, antiviral art-sRNAs are computationally designed to target one or multiple sites in viral RNAs with high specificity, and art-sRNA constructs are generated and introduced into plants that are subsequently challenged with the target virus(es). Numerous studies have reported the successful application of art-sRNAs to induce resistance against a large number of RNA and DNA viruses in model and crop species. However, the application of art-sRNAs as an antiviral tool has limitations, such as the difficulty to predict the efficacy of a particular art-sRNA or the emergence of virus variants with mutated target sites escaping to art-sRNA-mediated degradation. Here, we review the different classes, features, and uses of art-sRNA-based tools to induce antiviral resistance in plants. We also provide strategies for the rational design of antiviral art-sRNAs and discuss the latest advances in developing art-sRNA-based methodologies for enhanced resistance to plant viruses.
Highlights
RNA interference (RNAi) is a biological process conserved in most eukaryotes and characterized by the sequence-specific degradation of target RNA by complementary small RNAs [1].RNAi pathways are triggered by double-stranded RNA processed into sRNA duplexes byDicer ribonucleases [1,2]
The majority of artificial microRNAs (amiRNAs)-expressing plants were susceptible, and accumulated virus variants with mutated target sites [14]. These results suggest that the simultaneous multi-targeting of Tomato spotted wilt virus (TSWV) RNAs with various syn-tasiRNAs most likely limits the ability of the virus to mutate all target sites, whereas subinhibitory amiRNA accumulation favors the emergence of target site mutations in the replicating virus
Art-sRNAi tools have been broadly used in plants to confer antiviral resistance against multiple
Summary
RNA interference (RNAi) is a biological process conserved in most eukaryotes and characterized by the sequence-specific degradation of target RNA by complementary small RNAs (sRNAs) [1]. RNAi approaches, such as virus-induced gene silencing (VIGS) and hairpin (hp)-based silencing, consisted of the expression of dsRNA or hpRNA precursors, respectively, bearing sequences of the target virus (for a recent review see [5]). Very popular, these approaches lacked high specificity as the large populations of sRNAs produced from these type of precursors favor the accidental targeting of complementary cellular transcripts [6]. Despite differing in their biogenesis pathway (see below), both classes of art-sRNAs function by associating with AGO1 to silence target RNAs through endonucleolytic cleavage or translation repression (Figure 1)
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