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

Here we describe a robust sex-sorting approach termed SEPARATOR (Sexing Element Produced by Alternative RNA-splicing of A Transgenic Observable Reporter) that exploits sex-specific alternative splicing of an innocuous reporter to ensure exclusive dominant male-specific expression in Aedes aegypti. Using SEPARATOR, we demonstrate reliable sex sorting from larval and pupal stages, and use an Complex Object Parametric Analyzer and Sorter (COPAS) machine to demonstrate scalable high-throughput sex-selection of first instar larvae.