Search

Attraction and mating between male and female animals depend on effective communication between conspecifics. However, in mosquitoes, we have only a rudimentary understanding of the sensory cues and receptors critical for the communication that is essential for reproductive behavior. While it is known that male Aedes aegypti use sound to help them identify females, it is not unclear whether sound detection is absolutely required since other cues such as vision may also participate in mating behavior. To determine the effect of eliminating hearing on mating success, we knocked out the Ae. aegypti TRPVa channel, which is a protein expressed in chordotonal neurons in the Johnston's organ (JO) that respond to sound-induced movements in the antenna. Loss of trpVa eradicated sound-induced responses from the JO, thereby abolishing hearing. Strikingly, mutation of trpVa eliminated mating behavior in males. In contrast, trpVa -null females mated, although this behavior was slightly delayed relative to wild-type females. Males and females produce sounds as they beat their wings at distinct frequencies during flight. Sound mimicking the female wingbeat induced flight, attraction, and copulatory-like behavior in wild-type males without females present, but not in trpVa -null males. Males are known to modulate their wingbeat frequencies before mating in the air, which is a phenomenon referred to as rapid frequency modulation (RFM). We found that RFM was absent in mosquitoes lacking TRPVa. We conclude that the requirement for trpVa and hearing for male reproductive behavior in Aedes is absolute, as mating in the deaf males is eliminated., Competing Interests: Competing interests statement:The authors declare no competing interest.

In most experimental animals, it is challenging to combine mutations and rescue transgenes and to use bipartite systems to assess gene expression. To circumvent the difficulties in combining multiple genetic elements, we developed the DREaMR (Drug-on, REporter, Mutant, Rescue) system. Using Drosophila white as the initial model, we demonstrated that introduction of a single insertion by CRISPR/Cas9 created a null mutation, a tagged rescue construct, which could be induced with doxycycline, and which allowed assessment of protein expression. To create a DREaMR in an organism in which combining multiple genetic elements is more problematic than in Drosophila, we tested the mosquito, Aedes aegypti-the insect vector for dengue, yellow fever, Zika, and other viral diseases. We generated a DREaMR allele in the kh gene, which permitted us to induce expression of the rescue construct, and detect expression of Kh. Thus, this system avoids the need to perform genetic crosses to introduce an inducible rescue transgene in a mutant background, or to combine driver and reporter lines to examine expression of the targeted protein. We propose that DREaMR provides a system that can be applied to additional mosquito vectors as well as other organisms in which CRISPR/Cas9 is effective., (© The Author(s) 2021. Published by Oxford University Press on behalf of Genetics Society of America. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The mosquito Aedes aegypti is the principal vector for arboviruses including dengue/yellow fever, chikungunya, and Zika virus, infecting hundreds of millions of people annually. Unfortunately, traditional control methodologies are insufficient, so innovative control methods are needed. To complement existing measures, here we develop a molecular genetic control system termed precision-guided sterile insect technique (pgSIT) in Aedes aegypti. PgSIT uses a simple CRISPR-based approach to generate flightless females and sterile males that are deployable at any life stage. Supported by mathematical models, we empirically demonstrate that released pgSIT males can compete, suppress, and even eliminate mosquito populations. This platform technology could be used in the field, and adapted to many vectors, for controlling wild populations to curtail disease in a safe, confinable, and reversible manner., (© 2021. The Author(s).)

Aedes aegypti spread devastating viruses such as dengue, which causes disease among 100 to 400 million people annually. A potential approach to control mosquito disease vectors is the sterile insect technique (SIT). The strategy involves repeated release of large numbers of sterile males, which reduces insect populations because the sterile males mate and thereby suppress the fertility of females that would otherwise mate with fertile males. While SIT has been successful in suppressing certain agricultural pests, it has been less effective in depressing populations of Ae. aegypti This limitation is in part because of the fitness effects resulting from mutagenizing the mosquitoes nonspecifically. Here, we introduced and characterized the impact on female fertility of an Ae. aegypti mutation that disrupts a gene that is specifically expressed in testes. We used CRISPR/Cas9 to generate a null mutation in the Ae. aegypti β2-tubulin ( B2t ) gene, which eliminates male fertility. When we allowed wild-type females to first mate with B2t mutant males, most of the females did not produce progeny even after being subsequently exposed to wild-type males. We also introduced B2t mutant and wild-type males simultaneously with wild-type females and found that a larger number of B2t mutant males relative to the wild-type males was effective in significantly suppressing female fertility. These results raise the possibility of employing B2t sterile males to improve the efficacy of SIT in suppressing populations of Ae. aegypti through repeated releases and thereby reduce the transmission of viruses by these invasive mosquitoes., Competing Interests: The authors declare no competing interest.