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Generating cisgenic sexing strains in insect pests.

Serafima Davydova, Junru Liu, Nikolay P Kandul, Igor Antoshechkin, Jonathan Mann, W Evan Braswell, Omar S Akbari, Angela Meccariello

Communications biology · 2026-Feb-05 · PMID 41644661 · DOI 10.1038/s42003-026-09624-9

Abstract

Insect pest population control via sterile insect technique markedly benefits from separation by sex prior to release. To simplify this process, traditional genetics has been deployed to develop genetic sexing strains (GSSs) for several disease vectors and agricultural pests of vast economic significance, although very few are applied in the field due to associated fitness costs and instability. In this study, we generated a method to engineer cisgenic GSS (CGSS) in insects. We use CRISPR/Cas9-mediated homology-directed repair to seamlessly translocate a sex-specific alternatively spliced intron into a dominant phenotypic gene generating a genetically stable strain that enables sex-sorting by eye. To achieve this feat, we use Ceratitis capitata as our model and relied on the sex-specifically spliced intron of its endogenous transformer gene, which we seamlessly inserted a copy into the pupal colouration white pupae gene. This minimal modification resulted in the generation of a homozygous strain we term IMPERIAL that was genetically and phenotypically stable where all female pupae are brown while male pupae are white with overall good fitness. By minimally editing the genome, our novel CGSS approach can be applied to other pests that may aid more efficient and economically suitable pest control.

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Study type
laboratory
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published
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Conflict statement
Competing interests: The authors declare the following competing interests: A patent has been filed on this technology PCT/US2025/061221. O.S.A is a founder of Agragene, Inc. and Synvect, Inc. with equity interest. N.P.K is a founder of 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.

Topics and methods

Sterile insect techniquesSex determinationVector controlCRISPRSIT

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