Abstract
New mosquito control strategies are vitally needed to address established arthropod-borne infectious diseases such as dengue and yellow fever and emerging diseases such as Zika and chikungunya, all of which are transmitted by the disease vector mosquito Aedes aegypti. In this investigation, Saccharomyces cerevisiae (baker’s yeast) was engineered to produce short hairpin RNAs (shRNAs) corresponding to the Aedes aegypti orthologs of fasciculation and elongation protein zeta 2 (fez2) and leukocyte receptor cluster (lrc) member, two genes identified in a recent screen for A. aegypti larval lethal genes. Feeding A. aegypti with the engineered yeasts resulted in silenced target gene expression, disrupted neural development, and highly significant larval mortality. Larvicidal activities were retained following heat inactivation and drying of the yeast into tabular formulations that induced >95% mortality and were found to attract adult females to oviposit. These ready-to-use inactivated yeast interfering RNA tablets may one day facilitate the seamless integration of this new class of lure-and-kill species-specific biorational mosquito larvicides into integrated mosquito control programs.
Highlights
New mosquito control strategies are vitally needed to address established arthropod-borne infectious diseases such as dengue and yellow fever and emerging diseases such as Zika and chikungunya, all of which are transmitted by the disease vector mosquito Aedes aegypti
Two small interfering RNAs (siRNAs) molecules, #52 and #101, which correspond to the Aedes aegypti fasciculation and elongation zeta 2 and the receptor-encoding ortholog of leukocyte receptor cluster member genes, respectively, were identified in recent screens for mosquito larval lethal genes, which will be described in their entirety elsewhere
SiRNA #52, which corresponds to a target sequence in the first exon of fez[211] (Supplementary Fig. S1a), was identified in a chitosan nanoparticle larvicide screen in which it induced 40 ± 10% larval death (Fig. 1a; P = 0.00327 vs. control siRNA treatment)
Summary
Two siRNA molecules, #52 and #101, which correspond to the Aedes aegypti fasciculation and elongation zeta 2 (fez2) and the receptor-encoding ortholog of leukocyte receptor cluster (lrc) member genes, respectively, were identified in recent screens for mosquito larval lethal genes, which will be described in their entirety elsewhere These two interfering RNAs were prioritized, due to their high larvicidal activities and lack of conserved target sites in humans and other non-target organisms, for further characterization. S. cerevisiae is a source of nutrition and a strong odorant attractant that can act as a bait[5] to lure and kill A. aegypti larvae that hatch in the containers The results of this investigation indicate that genetically engineered yeast expressing interfering RNA corresponding to mosquito genes required for larval viability can function as lure-and-kill mosquito larvicides
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