Elimination of vision-guided target attraction in Aedes aegypti using CRISPR

Blood-feeding insects, such as the mosquito, Aedes (Ae.) aegypti, use multiple senses to seek out and bite humans.^1,2 Upon exposure to the odor of CO₂, the attention of female mosquitoes to potential targets is greatly increased. Female mosquitoes are attracted to high-contrast visual cues and use skin olfactory cues to assist them in homing in on targets several meters away.^3-9 Within close range, convective heat from skin and additional skin odors further assist the mosquitoes' evaluation as to whether the object of interest might be a host.^10,11 Here, using CRISPR-Cas9, we mutated the gene encoding Op1, which is the most abundant of the five rhodopsins expressed in the eyes of Ae. aegypti. Using cage and wind-tunnel assays, we found that elimination of op1 did not impair CO₂-induced target seeking. We then mutated op2, which encodes the rhodopsin most similar to Op1, and also found that there was no impact on this behavior. Rather, mutation of both op1 and op2 was required for abolishing vision-guided target attraction. In contrast, the double mutants exhibited normal phototaxis and odor-tracking responses. By measuring the walking optomotor response, we found that the double mutants still perceived optic flow. In further support of the conclusion that the double mutant is not blind, the animals retained an electrophysiological response to light, although it was diminished. This represents the first genetic perturbation of vision in mosquitoes and indicates that vision-guided target attraction by Ae. aegypti depends on two highly related rhodopsins.