Severson, D. W. | bioRxiv Science

bioRxiv2026

Soda-baited RNAi Yeast Insecticides as Effective Attractive Targeted Sugar Baits (ATSBs) for Mosquito Control

BackgroundAttractive Targeted Sugar Baits (ATSBs) are a promising, environmentally friendly approach for mosquito control, but the direct field application, scalability and long-term effectiveness of ATSBs across diverse mosquito species remain significant challenges. Methodology/Principal FindingsWe assessed the efficacy of a genetically engineered RNA interference (RNAi) yeast strain (Sh.463_56.10R) formulated in three sugar baits, soda (Coca-ColaTM), 10% sucrose, and a commercial mosquito bait (BaitStabTM), on Aedes, Anopheles and Culex mosquitoes. All RNAi yeast bait formulations induced significantly higher mean mortality (87-100%) compared to the control groups (0-9%; P<0.0001), but mosquitoes exhibited a higher feeding preference for RNAi yeast-soda baits, which induced mortality rates of 94-100% (P < 0.0001) recorded across all mosquito species. Additionally, to assess the competitiveness of the RNAi yeast-soda bait to other tropical sugar sources, semi-field choice assays were conducted in Trinidad, West Indies using competing flowering plants and fruits typically found in residential environments. The RNAi yeast-soda ATSB continued to perform well in the presence of competing floral and fruit sugar sources during both Aedes albopictus and Culex quiquefasciatus trials, though the presence of several fruits and flowers did reduce A. aegypti mortality, suggesting that further field testing will be necessary. The residual activity of the Sh.463_56.10R + soda formulation was retained for at least 14 months, with sustained 100% mortality in C. quinquefasciatus and 93-100% mortality in Aedes aegypti, Anopheles gambiae and Anopheles stephensi. The RNAi yeast-soda ATSB also performed well in semi-field studies performed with a prototype soda bottle feeder. Conclusions/SignificanceThis study demonstrates the potential of soda-baited RNAi yeast as a potent, long-lasting, and scalable platform for ATSB-based mosquito control as a component of integrated vector management programs. Author SummaryMosquito-borne diseases continue to affect millions of people worldwide, and current mosquito control methods face challenges such as low public uptake, insecticide resistance and environmental concerns. Here we evaluated a new and environmentally friendly approach to mosquito control using ATSBs. We tested genetically engineered species-specific yeast producing RNAi molecules capable of killing mosquitoes that feed on it. We mixed the yeast with three different sugar baits, including soda (Coca-ColaTM), 10% sucrose, and the commercial mosquito bait BaitStabTM formulation, and evaluated how well they worked against different mosquitoes. The results showed that the RNAi yeast mixed with soda was the most effective, killing up to 100% of mosquitoes in laboratory and outdoor tests. The bait remained effective in the presence of many competing natural tropical fruit and floral sugar sources. Remarkably, the bait, which can be delivered in a soda bottle feeder, stayed active for at least 14 months under simulated field conditions. These findings suggest that soda-based RNAi yeast baits could provide a practical, long-lasting and scalable tool for mosquito control and may help strengthen future strategies to reduce mosquito-borne diseases.

genetics↗

bioRxiv2021

Marker-assisted mapping enables effective forward genetic analysis in the arboviral vector Aedes aegypti, a species with vast recombination deserts

Aedes aegypti is a major vector of arboviruses that cause dengue, chikungunya, yellow fever and Zika. Although recent success in reverse genetics has facilitated rapid progress in basic and applied research, integration of forward genetics with modern technologies remains challenging in this important species, as up-to-47% of its chromosome is refractory to genetic mapping due to extremely low rate of recombination. Here we report the development of a marker-assisted-mapping (MAM) strategy to readily screen for and genotype only the rare but informative recombinants, drastically increasing both the resolution and signal-to-noise ratio. Using MAM, we mapped a transgene that was inserted in a >100 Mb recombination desert and a sex-linked spontaneous red-eye (re) mutation just outside the region. We subsequently determined, by CRISPR/Cas9-mediated knockout, that cardinal is the causal gene of re, which is the first forward genetic identification of a causal gene in Ae. aegypti. This study provides the molecular foundation for using gene-editing to develop versatile and stable genetic sexing methods by improving upon the current re-based genetic sexing strains. MAM does not require densely populated markers and can be readily applied throughout the genome to facilitate the mapping of genes responsible for insecticide- and viral-resistance. By enabling effective forward genetic analysis, MAM bridges a significant gap in establishing Ae. aegypti as a model system for research in vector biology. As large regions of suppressed recombination are also common in other plant and animal species including those of economic significance, MAM will have broad applications beyond vector biology.

genetics↗