Gene silencing by RNA interference in the house dust mite, Dermatophagoides pteronyssinus

Marr, Edward J; Sargison, Neil; Nisbet, Alasdair J.; Burgess, Stewart T. G.

doi:10.1016/j.mcp.2015.07.008

Cited by 17 publications

(23 citation statements)

References 38 publications

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“…We successfully established a suitable method to introduce dsRNA into S. scabiei , adapted from a previously published protocol to silence genes in house dust mites [37]. After 24 h incubation of mites at 4 °C with fluorescently labelled SsGST-mu1 dsRNA at 1.25 μg/μl, fluorescence was clearly visible in the mite gut (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…In some but not all mites dsRNA can be introduced simply and efficiently by feeding [52] which allows the dsRNA to enter cells and induce a systemic RNAi effect. Non-invasive immersion has been demonstrated to be the simplest and most gentle experimental method in the honey bee mite, V. destructor [33] and the house dust mite, D. pteronyssinus [37]. These mites have an experimental advantage as they are either truly ectoparasitic or free-living, respectively, whereas S. scabiei are obligatory living within host tissue and cannot be maintained in culture.…”

Section: Discussionmentioning

confidence: 99%

“…GSTs have also been associated with macrocyclic lactone resistance in T. urticae [40, 55] and S. scabiei [39]. We chose SsGST-mu1 as the first target to trial gene silencing in S. scabiei because this is one of the few S. scabiei genes that have been studied at the transcriptional level [39] and GSTs have been the target of choice in previously successful trials of RNAi in other mite species [33, 37]. GST s are housekeeping genes and have been found to be equally and constitutively expressed in synganglion, malpighian tubules and in the gut of the honey bee mite [33].…”

Section: Discussionmentioning

confidence: 99%

“…RNAi silencing has been demonstrated in a few mites, including the pest mites Varroa destructor (honey bee mite) [32, 33], Tetranychus urticae (two-spotted spider mite) [34] and Dermanysus gallinae (poultry red mite) [35] and also in Metaseiulus occidentalis [36], an agriculturally important biological control agent of plant-feeding pest mites. More recently RNAi has also been tested in Sarcoptiformes mites including the notorious house dust mite Dermatophagoides pteronyssinus [37] and the sheep scab mite, Psoroptes ovis (E. J. Marr personal communication). Glutathione S-transferase mu1 ( GST-mu1 ) has been the gene target of choice for most studies aiming to establish RNAi in mites.…”

Section: Introductionmentioning

confidence: 99%

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Gene silencing by RNA interference in Sarcoptes scabiei: a molecular tool to identify novel therapeutic targets

et al. 2017

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BackgroundScabies is one of the most common and widespread parasitic skin infections globally, affecting a large range of mammals including humans, yet the molecular biology of Sarcoptes scabiei is astonishingly understudied. Research has been hampered primarily due to the difficulty of sampling or culturing these obligatory parasitic mites. A further and major impediment to identify and functionally analyse potential therapeutic targets from the recently emerging molecular databases is the lack of appropriate molecular tools.MethodsWe performed standard BLAST based searches of the existing S. scabiei genome databases using sequences of genes described to be involved in RNA interference in Drosophila and the mite model organism Tetranychus urticae. Experimenting with the S. scabiei mu-class glutathione S-transferase (SsGST-mu1) as a candidate gene we explored the feasibility of gene knockdown in S. scabiei by double-stranded RNA-interference (dsRNAi).ResultsWe provide here an analysis of the existing S. scabiei draft genomes, confirming the presence of a double stranded RNA (dsRNA) - mediated silencing machinery. We report for the first time experimental gene silencing by RNA interference (RNAi) in S. scabiei. Non-invasive immersion of S. scabiei in dsRNA encoding an S. scabiei glutathione S-transferase mu-class 1 enzyme (SsGST-mu1) resulted in a 35% reduction in the transcription of the target gene compared to controls.ConclusionsA series of experiments identified the optimal conditions allowing systemic experimental RNAi without detrimental side effects on mite viability. This technique can now be used to address the key questions on the fundamental aspects of mite biology and pathogenesis, and to assess the potential therapeutic benefits of silencing S. scabiei target genes.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2226-1) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Gene silencing by RNA interference in Sarcoptes scabiei: a molecular tool to identify novel therapeutic targets

et al. 2017

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“…Reported expression knockdown in these experiments was between 70% and 91%, confirming efficient silencing. Also in other mites such as the house dust mite, Dermatophagoides pteronyssinus, the citrus red spider mite, Panonychus citri, and the red poultry mite, Dermanyssus gallinae, RNAi by feeding dsRNA was shown to be functional (Kamau et al, 2013;Marr et al, 2015;Tian et al, 2015b The present document has been produced and adopted by the bodies identified above as authors. This task has been carried out exclusively by the author(s) in the context of a contract between the European Food Safety Authority and the authors, awarded following a tender procedure.…”

Section: Cheliceratamentioning

confidence: 99%

Literature review of baseline information on RNAi to support the environmental risk assessment of RNAi‐based GM plants

Christiaens

Dzhambazova

Kostov

et al. 2018

EFS3

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This report is the outcome of an EFSA procurement (OC/EFSA/GMO/2015/02) which aims at reviewing relevant scientific information on RNA interference (RNAi) that could serve as baseline information for the environmental risk assessment of RNAi‐based genetically modified (GM) plants. The report is based on a systematic literature search on the use of RNAi molecules in arthropods, nematodes, annelids and molluscs with dsRNA, siRNA and miRNA applied primarily through feeding and soaking (oral ingestion). The numbers of retrieved publications covering these areas are reported, along with the species name, life stages tested, the target gene and its function, details of the test substances and their concentrations used, methods of delivery and effects. Separate sections discuss the available information on: (1) the uptake and systematic spread of RNAi activity, including a description of the various components involved in this process; (2) the mechanisms of dsRNA‐, siRNA‐ and miRNA‐elicited gene silencing and the different factors involved in RNAi efficiency; (3) routes of exposure of the biotic and abiotic environment to dsRNA, siRNA and miRNA from GM plants; (4) the environmental fate of dsRNA, siRNA and miRNA; and (5) the various factors that may limit non‐target effects including exposure, factors influencing the silencing efficiency of dsRNA, siRNA and miRNA, possible unintended and off‐target effects, and their mechanisms. Finally, an overview of the species of arthropods, nematodes, annelids and molluscs for which genomic data are available is also presented. The report identifies some of the challenges involved in developing plants with RNAi systems which affect invertebrate gene expression. The report also concludes that, currently, knowledge on issues such as exposure, specificity, offtarget effects, sequence similarities and bioinformatics is very limited, as only a few RNAi expressing plants which specifically target invertebrate species have been developed and comprehensively studied.

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Beyond insects: current status and achievements of RNA interference in mite pests and future perspectives

Niu

Shen

Christiaens

et al. 2018

Pest Management Science

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Mites comprise a group of key agricultural pests on a wide range of crops. They cause harm through feeding on the plant and transferring dangerous pathogens, and the rapid evolution of pesticide resistance in mites highlights the need for novel control methods. Currently, RNA interference (RNAi) shows great potential for insect pest control. Here, we review the literature regarding RNAi in mite pests. We discuss different target genes and RNAi efficiency in various mite species, a promising Varroa control program using RNAi, the synergy of RNAi with plant defense mechanisms and microorganisms, and current understanding of systemic movement of double-stranded RNA (dsRNA). On the basis of this evidence, we can conclude that there is clear potential for application of RNAi-based mite control, but further research on several aspects of RNAi in mites is needed, including: (i) the factors influencing RNAi efficiency, (ii) the mechanism of environmental RNAi and cross-kingdom dsRNA trafficking, (iii) the mechanism of possible systemic and parental RNAi, and (iv) non-target effects, specifically in predatory mites, which should be considered during RNAi target selection. © 2018 Society of Chemical Industry.

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Gene silencing by RNA interference in the house dust mite, Dermatophagoides pteronyssinus

Cited by 17 publications

References 38 publications

Gene silencing by RNA interference in Sarcoptes scabiei: a molecular tool to identify novel therapeutic targets

Gene silencing by RNA interference in Sarcoptes scabiei: a molecular tool to identify novel therapeutic targets

Literature review of baseline information on RNAi to support the environmental risk assessment of RNAi‐based GM plants

Beyond insects: current status and achievements of RNA interference in mite pests and future perspectives

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