Gene silencing in adult <i>Aedes aegypti</i> mosquitoes through oral delivery of double‐stranded RNA

Coy, Monique R.; Sanscrainte, Neil D.; Chalaire, K. C.; Inberg, Alex; Maayan, Inbar; Glick, Eitan; Paldi, Nitzan; Becnel, James J.

doi:10.1111/j.1439-0418.2012.01713.x

Cited by 51 publications

(48 citation statements)

References 52 publications

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“…Although ingestion-based strategies do not work in all insect species, perhaps most notably Drosophila melanogaster , oral delivery of interfering RNA mixed with food has promoted gene silencing in a variety of insects 8,17 , including A. aegypti adults 18 . We described chitosan nanoparticle-mediated RNAi in A. gambiae larvae 19 and have successfully applied this approach for reduction of gene expression in A. aegypti larvae 20,21 .…”

Section: Introductionmentioning

confidence: 99%

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Zhang

Mysore

Flannery

et al. 2015

JoVE

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SHORT ABSTRACT Here we describe a procedure for inhibiting gene function in disease vector mosquitoes through the use of chitosan/interfering RNA nanoparticles that are ingested by larvae. LONG ABSTRACT Vector mosquitoes inflict more human suffering than any other organism—and kill more than one million people each year. The mosquito genome projects facilitated research in new facets of mosquito biology, including functional genetic studies in the primary African malaria vector Anopheles gambiae and the dengue and yellow fever vector Aedes aegypti. RNA interference- (RNAi-) mediated gene silencing has been used to target genes of interest in both of these disease vector mosquito species. Here, we describe a procedure for preparation of chitosan/interfering RNA nanoparticles that are combined with food and ingested by larvae. This technically straightforward, high-throughput, and relatively inexpensive methodology, which is compatible with long double stranded RNA (dsRNA) or small interfering RNA (siRNA) molecules, has been used for the successful knockdown of a number of different genes in A. gambiae and A. aegypti larvae. Following larval feedings, knockdown, which is verified through qRT-PCR or in situ hybridization, can persist at least through the late pupal stage. This methodology may be applicable to a wide variety of mosquito and other insect species, including agricultural pests, as well as other non-model organisms. In addition to its utility in the research laboratory, in the future, chitosan, an inexpensive, non-toxic and biodegradable polymer, could potentially be utilized in the field.

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Section: Introductionmentioning

confidence: 99%

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Zhang

Mysore

Flannery

et al. 2015

JoVE

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“…Therefore the reduction in virus may be due to an overall impact on mosquito health following V-ATPase silencing. Conversely, Coy et al (2012) showed that dsRNA delivered orally suppresses V-ATPase subunit C by 60% (168 h post exposure) without any noticeable death [11]. Death may have not been observed by Coy et al (2012) because animals were monitored only for 48 h following exposure (as compared to 50 days observed by Kang et al (2014)) [11,80].…”

Section: Rnai Triggers With Potential Mosquito Control Applicationsmentioning

confidence: 99%

“…This was noted in Ae. aegypti where vATPase C dsRNA was found partially degraded in tissues after 24 h [11]. The transcript was reduced by 60% but failed to kill the mosquito.…”

Section: Delivery Systems For Rnai Triggers In Vivomentioning

confidence: 99%

“…With hundreds of documented effective RNAi triggers targeting mosquito and pathogen genes, there is an expansive arsenal of anti-vector and anti-pathogen targets that could be harnessed for mosquito and mosquito-borne disease control strategies. RNAi experiments in a number of Aedes , Anopheles , Culex , and Armigeres disease vector species have resulted in disruption of processes including: morphogenesis, olfaction for host seeking and oviposition, blood feeding, fertility, fecundity, and survival [10,11,12,13,14,15,16,17,18,19]. …”

Section: Introductionmentioning

confidence: 99%

“…Additionally, strategically placed ATSB stations near breeding sites (dubbed Attractive Baited Oviposition Trap, ABOT) or indoors can attract and kill vector species in proximity to people [32,33,40,41]. Although ATSB have not been studied in conjunction with RNAi, successful gene silencing by oral exposure routes has been documented using sucrose meals and artificial blood meals demonstrating the potential in combining these control approaches [11,42]. By the same logic, mosquitocidal RNAi triggers could be applied to target essential genes for embryogenesis in Attractive Baited Oviposition Traps (ABOTs) [43,44,45].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

RNA Interference for Mosquito and Mosquito-Borne Disease Control

Airs

Bartholomay

2017

Insects

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RNA interference (RNAi) is a powerful tool to silence endogenous mosquito and mosquito-borne pathogen genes in vivo. As the number of studies utilizing RNAi in basic research grows, so too does the arsenal of physiological targets that can be developed into products that interrupt mosquito life cycles and behaviors and, thereby, relieve the burden of mosquitoes on human health and well-being. As this technology becomes more viable for use in beneficial and pest insect management in agricultural settings, it is exciting to consider its role in public health entomology. Existing and burgeoning strategies for insecticide delivery could be adapted to function as RNAi trigger delivery systems and thereby expedite transformation of RNAi from the lab to the field for mosquito control. Taken together, development of RNAi-based vector and pathogen management techniques & strategies are within reach. That said, tools for successful RNAi design, studies exploring RNAi in the context of vector control, and studies demonstrating field efficacy of RNAi trigger delivery have yet to be honed and/or developed for mosquito control.

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Delivery of gene‐specific dsRNA by microinjection and feeding induces RNAi response in Sri Lanka weevil, Myllocerus undecimpustulatus undatus Marshall

Pinheiro

Taylor

et al. 2019

Pest Management Science

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BACKGROUND RNA interference (RNAi) is widely used in entomological research for functional analysis of genes and is being considered as a new tool for insect pest management. Sri Lanka weevil (SLW) is a highly polyphagous pest of agronomically important plants, but currently only a few control methods are available for this insect. RESULTS In the present study, we evaluated the stability of candidate reference genes β‐ACT, α‐TUB, EF1‐α, RPL12 and GAPDH, and identified EF1‐α as the most reliable for gene expression normalization. Four target genes involved in different cellular processes, including Prosα2, RPS13, Snf7 and V‐ATPase A were selected to evaluate whether RNAi response in SLW adults can be triggered by microinjection and oral feeding of their double‐stranded RNAs (dsRNAs). Three days after injection of the dsRNAs for the target genes, their transcript levels were significantly reduced (up to 91.4%) when compared to the control. Additionally, weevils fed with the target dsRNAs showed significant decreases in gene transcript levels and significant mortality was observed in insects treated with Prosα2 and Snf7 dsRNAs (78.6 to 92.7%). CONCLUSION Our data demonstrate that microinjection and feeding of dsRNA produce a strong RNAi response in SLW, indicating that RNAi‐based strategies could be explored to develop a selective and environmentally safe control method against SLW. © 2019 Society of Chemical Industry

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Gene silencing in adult Aedes aegypti mosquitoes through oral delivery of double‐stranded RNA

Cited by 51 publications

References 52 publications

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

RNA Interference for Mosquito and Mosquito-Borne Disease Control

Delivery of gene‐specific dsRNA by microinjection and feeding induces RNAi response in Sri Lanka weevil, Myllocerus undecimpustulatus undatus Marshall

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