Efficient sex separation by exploiting differential alternative splicing of a dominant marker in Aedes aegypti.
Shih-Che Weng, Igor Antoshechkin, Eric Marois, Omar S Akbari
PLoS genetics · 2023-Nov-27 · PMID 38011259 · DOI 10.1371/journal.pgen.1011065Abstract
Only female mosquitoes consume blood giving them the opportunity to 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 (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. Using SEPARATOR, we demonstrate reliable sex selection from early 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. 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.
Evidence profile
- Study type
- laboratory
- Text reviewed by the copilot
- Full text is available
- Publication status
- published
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- Conflict statement
- I have read the journal’s policy and the authors of this manuscript have the following competing interests: O.S.A. and S.W. have filed a patent on this technology. O.S.A is a founder of Agragene, Inc. and Synvect, Inc. with equity interest. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. All other authors declare no competing interests.