Abstract
Aedes aegypti is a vector of dengue, chikungunya, and Zika viruses. Current vector control strategies such as community engagement, source reduction, and insecticides have not been sufficient to prevent viral outbreaks. Thus, interest in novel strategies involving genetic engineering is growing. Female mosquitoes rely on flight to mate with males and obtain a bloodmeal from a host. We hypothesized that knockout of genes specifically expressed in female mosquitoes associated with the indirect flight muscles would result in a flightless female mosquito. Using CRISPR-Cas9 we generated loss-of-function mutations in several genes hypothesized to control flight in mosquitoes, including actin (AeAct-4) and myosin (myo-fem) genes expressed specifically in the female flight muscle. Genetic knockout of these genes resulted in 100% flightless females, with homozygous males able to fly, mate, and produce offspring, albeit at a reduced rate when compared to wild type males. Interestingly, we found that while AeAct-4 was haplosufficient, with most heterozygous individuals capable of flight, this was not the case for myo-fem, where about half of individuals carrying only one intact copy could not fly. These findings lay the groundwork for developing novel mechanisms of controlling Ae. aegypti populations, and our results suggest that this mechanism could be applicable to other vector species of mosquito.
Highlights
The yellow fever mosquito Aedes aegypti is a vector for many viruses of medical significance, such as dengue, Zika, chikungunya, and yellow fever, and they can be found in tropical, subtropical, and temperate regions of the world [1]
We identify several genes that are critical only for female mosquito flight; genetic ablation of each gene resulted in flightless females, but flying males
Due to a lack of safe, effective vaccines for most viruses transmitted by Ae. aegypti, save yellow fever virus [4], control efforts largely focus on reducing vector abundance, and include source reduction, and chemical methods like insecticides or larvicides [3,5]
Summary
The yellow fever mosquito Aedes aegypti is a vector for many viruses of medical significance, such as dengue, Zika, chikungunya, and yellow fever, and they can be found in tropical, subtropical, and temperate regions of the world [1]. After hatching from the embryo, Ae. aegypti like all other mosquito species will progress through the aquatic larval and pupal stages of their life, before emerging as an adult from the pupal casing to fly away [2,3]. Due to a lack of safe, effective vaccines for most viruses transmitted by Ae. aegypti, save yellow fever virus [4], control efforts largely focus on reducing vector abundance, and include source reduction, and chemical methods like insecticides or larvicides [3,5]. Additional concerns relating to the emergence of resistance and effects on off-target species are increasing [6]. Because of these limitations, the need for novel vector control strategies is growing
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