A dsRNA‐degrading nuclease (dsRNase2) limits RNAi efficiency in the Asian corn borer (<i>Ostrinia furnacalis</i>)

Fan, Yunhe; Song, Huifang; Abbas, Mureed; Wang, Yanli; Li, Tao; Ma, Enbo; Cooper, Anastasia M.W.; Silver, Kristopher; Zhu, Kun Yan; Zhang, Jianzhen

doi:10.1111/1744-7917.12882

Cited by 32 publications

(41 citation statements)

References 51 publications

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“…To improve the knockdown efficiency, dsRNA of dsRNase gene is sequentially‐ or co‐fed with a target RNAi gene as a way of enhancing dsRNA stability in insect (Chung et al, 2018; Fan et al, 2020; Garcia et al, 2017; Luo et al, 2017; Peng, Wang, Chen, et al, 2020; Prentice et al, 2019; Song et al, 2017; Spit et al, 2017; Tayler et al, 2019; Yoon et al, 2020). Moreover, the encapsulation of naked dsRNA has been used for improving dsRNA delivery and protecting dsRNA degradation from dsRNases.…”

Section: Discussionmentioning

confidence: 99%

“…RNAi is successful in some coleopteran insects, whereas it is relatively ineffective in other orders (Miller et al, 2012; Terenius et al, 2011; Zhu et al, 2011). The major reason is the presence of dsRNases in saliva, gut, or hemolymph that induces the degradation of introduced dsRNAs (Allen & Walker, 2012; Arimatsu et al, 2007; Cooper et al, 2020; Fan et al, 2020; Garcia et al, 2017; Luo et al, 2017; Peng, Wang, Zhu, et al, 2020; Prentice et al, 2019; Shukla et al, 2016; Song et al, 2017; Spit et al, 2017; Wang et al, 2016; Wynant et al, 2014). Another reason is the entrapment of dsRNAs in endosomes after endocytosis, resulting in an inefficient RNAi response, as shown in a lepidopteran cell line (Yoon et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

“…One is the encapsulation of dsRNAs using for examples lipoproteins, nanoparticles, or exosomes, which can protect dsRNA from degradation by dsRNase and increase the intracellular transportation (Das et al, 2015; He et al, 2013; Zhang et al, 2010, 2015; Zheng et al, 2019; Zhu & Palli, 2020). The other is the co‐feeding or sequential feeding with dsRNA of dsRNase genes, which improves the knockdown efficiency by increasing the target dsRNA stability (Chung et al, 2018; Fan et al, 2020; Garcia et al, 2017; Luo et al, 2017; Peng, Wang, Chen, et al, 2020; Prentice et al, 2019; Song et al, 2017; Spit et al, 2017; Tayler et al, 2019; Yoon et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

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Identification and functional analysis of dsRNases in spotted‐wing drosophila, Drosophila suzukii

Yoon

Ahn

Flinn

et al. 2021

Arch Insect Biochem Physiol

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RNAi efficiency in insects is different from species to species; some species in Coleoptera are relatively more amenable to RNA interference (RNAi) than other species.One of the major factors is the presence of dsRNAdegrading enzymes, called dsRNases, in saliva, gut, or hemolymph in insects, which degrade the double-stranded RNA (dsRNA) introduced, resulting in the low efficacy of RNAi. In this study, we report a dsRNA-degrading activity in the gut homogenates from the spotted-wing drosophila, Drosophila suzukii, by ex vivo assay. Then, we identified two Drosophila suzukii dsRNase genes, named DrosudsRNase1 and DrosudsRNase2. In silico analysis shows that the gene structures are similar to dsRNases found in other insects.When dsRNases expressed in Sf9 cells were compared for their dsRNA degrading activities, dsRNase1 was more vital than dsRNase2. Both dsRNases were expressed highly and exclusively in the gut compared to the rest of body. Also, they were highly expressed during larval and adult stages but not in embryonic and pupal stages, suggesting the dsRNases protect foreign RNA molecules received during the feeding periods. DsRNase1 was expressed at a higher level in adults, whereas dsRNase2 showed more expression in early larvae. Our study on the tissue and development-This article has been contributed to by US Government employees and their work is in the public domain in the USA.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Identification and functional analysis of dsRNases in spotted‐wing drosophila, Drosophila suzukii

Yoon

Ahn

Flinn

et al. 2021

Arch Insect Biochem Physiol

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“…In vivo co-targeting of dsRNases together with other vital genes has been demonstrated to significantly reduce gene expression and insect performance, compared to targeting only the vital gene, in insects of the orders Coleoptera (Almeida Garcia et al, 2017;Spit et al, 2017;Prentice et al, 2019;Peng et al, 2020a), Hemiptera (Luo et al, 2017;Chung et al, 2018;Kaur et al, 2020;Sharma et al, 2021), and Diptera (Tayler et al, 2019;Giesbrecht et al, 2020). Additional studies also suggest that inadequate oral RNAi is due to the presence of dsRNases in an even greater diversity of insect taxa (Song et al, 2017;Ghodke et al, 2019;Cooper et al, 2020;Fan et al, 2020;Peng et al, 2020b;Yoon et al, 2021).…”

Section: Targeting Dsrnasesmentioning

confidence: 99%

RNAi Targets in Agricultural Pest Insects: Advancements, Knowledge Gaps, and IPM

Willow¹,

Taning²,

Cook³

et al. 2021

Front. Agron.

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The unprecedented target-specificity of double-stranded RNA (dsRNA), due to its sequence-specific mode of action, puts dsRNA at the forefront of biosafe insecticide technology. Since 2007, sensitive target genes have been identified in numerous crop pest insects, with an end goal of applying RNA interference (RNAi) in pest management. Key RNAi targets identified include genes involved in (1) feeding and digestion, (2) production of dsRNases, (3) resistance to insecticides and plant allelochemicals, (4) reproductive fitness, and (5) transmission of plant viruses. Despite the advances, there remain critical knowledge gaps in each of these areas. Particular emphasis must be placed on ensuring RNAi's compatibility with integrated pest management (IPM), via further identification of molecular targets that reduce crop damage while sustaining pest (host) populations for highly specialized biocontrol agents, the latter representing a core pillar of IPM.

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“…Recent studies have demonstrated that degradation of dsRNA by dsRNAdegrading nucleases (dsRNase) may occur prior to cellular uptake. Extracellular degradation of dsRNA, notably in the gut, may explain the low efficiency of oral RNAi (Garbutt et al, 2013;Singh et al, 2017;Peng et al, 2018;Prentice et al, 2019;Cooper et al, 2020;Fan et al, 2020;Chen et al, 2021;Yoon et al, 2021). The first insect dsRNase identified in the midgut fluid of the domestic silk moth (Bombyx mori), contains a signal peptide and a non-specific endonuclease domain (Arimatsu et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Cloning and Functional Characterization of a Double-Stranded RNA-Degrading Nuclease in the Tawny Crazy Ant (Nylanderia fulva)

et al. 2022

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RNA interference is a powerful tool that post-transcriptionally silences target genes. However, silencing efficacy varies greatly among different insect species. Recently, we attempted to knock down some housekeeping genes in the tawny crazy ant (Nylanderia fulva), a relatively new invasive species in the southern United States, but only achieved relatively low silencing efficiency when dsRNA was orally administered. Here, we detected divalent cation-dependent, dsRNA-degrading activity in the midgut fluid of worker ants in ex vivo assays. To determine whether dsRNA degradation could contribute to low effectiveness of oral RNAi in N. fulva, we cloned its sole dsRNase gene (NfdsRNase). The deduced amino acid sequence contained a signal peptide and an endonuclease domain. Sequence alignment indicated a high degree of similarity with well-characterized dsRNases, particularly the six key residues at active sites. We also identified dsRNase homologs from five other ant species and found a tight phylogenetic relationship among ant dsRNases. NfdsRNase is expressed predominantly in the abdomen of worker ants. Oral delivery of dsRNA of NfdsRNase significantly reduced the expression of NfdsRNase transcripts, and substantially suppressed dsRNA-degrading activity of worker ants’ midgut fluids as well. Our data suggest that dsRNA stability in the alimentary tract is an important factor for gene silencing efficiency in N. fulva, and that blocking NfdsRNase in gut lumen could potentially improve RNAi, a novel pest management tactic in control of N. fulva and other ant species.

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A dsRNA‐degrading nuclease (dsRNase2) limits RNAi efficiency in the Asian corn borer (Ostrinia furnacalis)

Cited by 32 publications

References 51 publications

Identification and functional analysis of dsRNases in spotted‐wing drosophila, Drosophila suzukii

Identification and functional analysis of dsRNases in spotted‐wing drosophila, Drosophila suzukii

RNAi Targets in Agricultural Pest Insects: Advancements, Knowledge Gaps, and IPM

Cloning and Functional Characterization of a Double-Stranded RNA-Degrading Nuclease in the Tawny Crazy Ant (Nylanderia fulva)

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