Improving the sex-specificity of a conditional female lethal system for genetic biocontrol of the New World screwworm, Cochliomyia hominivorax.

The New World screwworm is an obligate parasitic fly and a significant economic pest of livestock in the Americas. Although eradicated from the USA using the Sterile Insect Technique (SIT), enhancing SIT efficiency remains a priority. A promising approach involves conditional female-lethal genetic strains that produce only males in the absence of tetracycline, ideally eliminating females early in development to reduce larval diet costs. However, while some strains match wild-type production levels, lower male fitness reduces the net benefit of replacing the current wild-type strain with one of these genetic-sexing strains. This study aimed to improve strain performance through female-specific expression of both the driver and effector components of the lethality system. We tested four transgenic strains using early embryo-specific promoters from the Chhalo and g6451 genes. Strains with the Chhalo promoter driving tTA expression exhibited early-stage female lethality under a modified doxycycline regimen but suffered from reduced male fitness. In contrast, one strain with the g6451 promoter produced males with excellent fitness but female lethality occurred at the late pupal stage. Despite imperfect female lethality timing, the overall fitness characteristics of this strain makes it a good candidate for future sterile or fertile male release genetic control programs.