A Yeast RNA-Interference Pesticide Targeting the Irx Gene Functions as a Broad-Based Mosquito Larvicide and Adulticide

SIMPLE SUMMARY: It is critical that we identify new methods of preventing mosquito-borne infectious diseases, which threaten millions of people worldwide. In this investigation, we describe characterization of a new insecticide that turns off the mosquito Iroquois (Irx) gene, which is required for mosquito survival. The pesticide is synthesized in yeast, which can be fed to adult mosquitoes in a sugar bait solution or to juvenile mosquitoes that eat the yeast when it is placed in water where mosquitoes breed. Although the yeast kills several different types of mosquitoes, it was not found to affect the survival of other types of arthropods that consumed the yeast. These results indicate that yeast insecticides could one day be used for environmentally friendly mosquito control and disease prevention. ABSTRACT: Concerns for widespread insecticide resistance and the unintended impacts of insecticides on nontarget organisms have generated a pressing need for mosquito control innovations. A yeast RNAi-based insecticide that targets a conserved site in mosquito Irx family genes, but which has not yet been identified in the genomes of nontarget organisms, was developed and characterized. Saccharomyces cerevisiae constructed to express short hairpin RNA (shRNA) matching the target site induced significant Aedes aegypti larval death in both lab trials and outdoor semi-field evaluations. The yeast also induced high levels of mortality in adult females, which readily consumed yeast incorporated into an attractive targeted sugar bait (ATSB) during simulated field trials. A conserved requirement for Irx function as a regulator of proneural gene expression was observed in the mosquito brain, suggesting a possible mode of action. The larvicidal and adulticidal properties of the yeast were also verified in Aedes albopictus, Anopheles gambiae, and Culex quinquefasciatus mosquitoes, but the yeast larvicide was not toxic to other nontarget arthropods. These results indicate that further development and evaluation of this technology as an ecofriendly control intervention is warranted, and that ATSBs, an emerging mosquito control paradigm, could potentially be enriched through the use of yeast-based RNAi technology.