Efficient Sex Separation by Exploiting Differential Alternative Splicing of a Dominant Marker in Aedes aegypti
Weng S, Antoshechkin I, Marois E, Akbari OS.
10.1101/2023.06.16.545348Abstract
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®) to demonstrate scalable high-throughput sex-selection of first instar larvae. Additionally, we use this approach to sequence the transcriptomes of early larval males and females and find several genes that are sex-specifically expressed in males. SEPARATOR can simplify mass production of males for release programs and is designed to be cross-species portable and should be instrumental for genetic biocontrol interventions.
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Efficient sex separation by exploiting differential alternative splicing of a dominant marker in Aedes aegypti. · DOI 10.1371/journal.pgen.1011065