Efficient Sex Separation by Exploiting Differential Alternative Splicing of a Dominant Marker in Aedes aegypti .

Only female mosquitoes consume blood and transmit deadly human pathogens. Therefore, it is critical to remove females before conducting releases for genetic biocontrol interventions. Here we describe a robust sex-sorting approach termed SEPARATOR ( S exing E lement P roduced by A lternative R NA-splicing of A T ransgenic O bservable R eporter) that exploits sex-specific alternative splicing of an innocuous reporter to ensure exclusive dominant male-specific expression. Using SEPARATOR, we demonstrate reliable sex selection from larval and pupal stages in Aedes aegypti , and use a Complex Object Parametric Analyzer and Sorter (COPAS®) machine to demonstrate scalable high-throughput sex-selection of first instar larvae and beyond. SEPARATOR can simplify mass production of males for release approaches and is designed to be cross-species portable for streamlined development in other species and should be instrumental for genetic biocontrol interventions.