RNA interference in the cat flea, Ctenocephalides felis: Approaches for sustained gene knockdown and evidence of involvement of Dicer-2 and Argonaute2

Edwards, Catriona H; Baird, John; Zinser, Erich W.; Woods, Debra J.; Shaw, Sophie; Campbell, Ewan M.; Bowman, Alan K.

doi:10.1016/j.ijpara.2018.04.006

Cited by 6 publications

(4 citation statements)

References 50 publications

(59 reference statements)

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“…RNAi has proven an effective means to silence targeted genes in a range of pest arthropod species 3,23,36 . This has allowed more detailed research into physiological pathways and molecular mechanisms such as reproductive behaviour and immune response 37,38 .…”

Section: Discussionmentioning

confidence: 99%

Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum

Edwards

Christie

Masotti

et al. 2020

Sci Rep

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In this study, the use of dendrimer-coated carbon nanotubes (CNTs) as a delivery vehicle for dsRNA was assessed in Tribolium castaneum. Exposure to low dosages of polyamidoamine dendrimer carbon nanotubes (PAMAM-CNTs) did not affect T. castaneum larval mortality. Expression of key apoptotic factors, Dronc (Tc12580), Dredd (Tcn-like, Tc014026) and Buffy, (Tcinhib apop1), which can act as toxicity indicators, were not altered in T. castaneum larvae following injection of PAMAM-CNTs. The level of knockdown of two target genes, α-tubulin and mitochondrial RNA polymerase (mtpol), were significantly increased when larvae were injected with double-stranded RNA bound to CNTs (PAMAM-CNT-dsRNA), compared to those injected with target dsRNA alone. PAMAM-CNTs were visualised in cellular vacuoles and in the cell nucleus. Increase occurrence of a blistered wing phenotype was found in a subset of PAMAM-CNT-dsRNA αtub injected larvae, relative to the level seen in larvae injected with naked dsRnA αtub alone. These results suggest that the use of functionalised CNTs for dsRNA delivery could increase the efficacy of RNA interference in insect pest species. RNA interference (RNAi) is a promising tool for the control of insect pests. Introduction of exogenous doublestranded RNA (dsRNA) is effective at triggering gene knockdown and associated phenotypes in many problem pest species such as the Western Corn Rootworm 1 , the Colorado potato beetle 2 and the Varroa mite 3. New developments in dsRNA delivery methods to crop pests, such as dsRNA expression in transgenic plants 4 and foliar application 5 , offer the possibility of dsRNA-based insecticides 6. Despite this promise, there is notable variation in RNAi response between insect species 7 , with some, such as many lepidopterans 8 , unable to mount a strong RNAi response to dsRNA. Ingestion of dsRNA can be a valid route for delivery, yet many insects secrete enzymes in their gut that breakdown dsRNA before the latter can elicit an effect on target gene transcripts 9-11. When dsRNA enters into the cytoplasm, it is generally very effective in mounting an RNAi response 12,13. However, the route to the cytoplasm can act as a barrier to effective RNAi, with stability and uptake of dsRNA within the insect thought to be the significant limiting factor 10,11,14,15. Therefore, the protection of dsRNA from degradation coupled with an efficient cellular uptake is key to RNAi success. The development of delivery vehicles or carriers that enable these two processes is now necessary to fully realise the potential for dsRNA as an effective control measure. Combining dsRNA with other substances that act as efficient carriers, such as chitosan 16,17 , perfluorocarbon nanoparticles 18-20 or ribonuceloparticles 21 , can stabilise dsRNA and increase the chances of successful gene knockdown. Encapsulation of dsRNA within carbon quantum dots 22 and liposomes 23,24 has also shown some promise in increasing efficacy in more challenging insect species. In the last decade, carbon nanotubes (CNTs) have emerg...

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Section: Discussionmentioning

confidence: 99%

Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum

Edwards

Christie

Masotti

et al. 2020

Sci Rep

Self Cite

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“…While injection constitutes a reliable system to confirm as proof of concept the efficiency of gene silencing, dsRNA-based biopesticides need to work efficiently upon ingestion or contact in field conditions. Activity of digestive enzymes however may lead to degradation of dsRNA molecules in some insects, though for other species diets with addition of dsRNA showed a strong silencing effect (e.g., Ctenocephalides felis Bouché (Siphonaptera: Pulicidae), Edwards et al, 2018). A fundamental aspect for the activity of RNAi is the ability of the dsRNA molecules to pass cellular membranes in order to allow the uptake of exogenous dsRNA in the cells of the target organism and its subsequent spread.…”

Section: Rnai: Mode Of Action Advantages and Limitationsmentioning

confidence: 99%

The use of RNA interference for the management of arthropod pests in livestock farms

Bonina,

Arpaia

2023

Medical Vet Entomology

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Pest management in farm animals is an important action to contain economic damage to livestock production and prevent transmission of severe diseases to the stock. The use of chemical insecticides is still the most common approach followed by farmers; however, avoiding possible toxic effects on animals is a fundamental task for pest control measures compatible with animal well‐being. Moreover, legal constraints and insurgence of resistance by target species to the available insecticidal compounds are increasingly complicating farmers' operations. Alternatives to chemical pesticides have been explored with some promising results in the area of biological control or the use of natural products as sprays. The application of RNA interference techniques has enabled the production of new means of pest control in agriculture, and it is opening a promising avenue for controlling arthropod pests of livestock. Transcript depletion of specific target genes of the recipient organisms is based on the action of double‐strand RNAs (dsRNA) capable of impairing the production of fundamental proteins. Their mode of action, based on the specific recognition of short genomic sequences, is expected to be highly selective towards non‐target organisms potentially exposed; in addition, there are physical and chemical barriers to dsRNA uptake by mammalian cells that render these products practically innocuous for higher animals. Summarising existing literature on gene silencing for main taxa of arthropod pests of livestock (Acarina, Diptera, Blattoidea), this review explores the perspectives of practical applications of dsRNA‐based pesticides against the main pests of farm animals. Knowledge gaps are summarised to stimulate additional research in this area.

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“…The delivery system allows for the knockdown of very specific targeted gene expression in both insects and acarines. Edwards et al [ 128 ] were able to demonstrate the transfer of RNAi to cat fleas through a membrane feeding system, and resulted in 96% knockdown of GSTσ, a detoxification enzyme, within 2 days and sustained at least 7 days. The RNAi response to ingested dsRNA in C. f. felis was not impaired by gut enzymes of the flea.…”

Section: Future Directionsmentioning

confidence: 99%

Recent Advancements in the Control of Cat Fleas

Rust¹

2020

Insects

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With the advent of imidacloprid and fipronil spot-on treatments and the oral ingestion of lufenuron, the strategies and methods to control cat fleas dramatically changed during the last 25 years. New innovations and new chemistries have highlighted this progress. Control strategies are no longer based on the tripartite approach of treating the pet, the indoor environment, and outdoors. The ability of modern therapies to break the cat flea life cycle and prevent reproduction has allowed for the stand-alone treatments that are applied or given to the pet. In doing so, we have not only controlled the cat flea, but we have prevented or reduced the impact of many of the diseases associated with ectoparasites and endoparasites of cats and dogs. This review provides an update of newer and non-conventional approaches to control cat fleas.

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RNA interference in the cat flea, Ctenocephalides felis: Approaches for sustained gene knockdown and evidence of involvement of Dicer-2 and Argonaute2

Abstract: Graphical abstract

Cited by 6 publications

References 50 publications

Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum

Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum

The use of RNA interference for the management of arthropod pests in livestock farms

Recent Advancements in the Control of Cat Fleas

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