The sex determination gene doublesex is essential for female-specific flight and hearing properties in Aedes aegypti mosquitoes

The acoustic communication system supporting reproduction in disease-transmitting mosquitoes is sexually dimorphic, from the sounds that mosquitoes make to the properties of their flagellar ears. The molecular mechanisms shaping these dimorphisms, however, remain unclear. Here, we explored the contributions of the female-specific isoform of the sex determination gene doublesex (dsxF) in specifying female-unique properties in the acoustic communication system of Aedes aegypti mosquitoes. CRISPR-Cas9 based generation of a novel dsxF knockout line found that dsxF mutant females were flightless due to lack of expression of a female-specific myosin gene, myo-fem, in the thorax. We found that DsxF directly binds to four different enhancer sites of myo-fem to promote gene expression. We observed that dsxF female ears exhibited intersex neuroanatomical characteristics and contained a more extensive auditory efferent network than control females. At the peripheral hearing function level, dsxF female ears displayed intersex tuning frequencies associated with increased expression of hearing genes in dsxF female ears compared to control females. We identified three hearing genes transcriptionally inhibited by DsxF via direct binding. DsxF is a bifunctional transcription factor, activating female-specific gene expression in thoraces whilst repressing male-biased gene expression in ears to coordinate female-unique features across sensory and motor systems.