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<p>Effective sex separation remains a critical challenge for mosquito genetic control. Genetic sexing strains (GSS) address this by linking maleness with selectable traits, enabling efficient female removal. Here, we present a versatile platform for GSS development in the invasive <i>Aedes albopictus</i> mosquito that integrates CRISPR-engineered selectable phenotypes with sex conversion via <i>nix</i>, the male-determining factor. As a proof-of-concept, we disrupt the <i>yellow</i> pigmentation gene and restore its function in males using <i>nix</i>-containing transgenes, producing a stable strain with <i>yellow</i> females and dark males. Beyond serving as a vivid marker, <i>yellow</i> confers added advantages: GSS females pupate later than wild females, enhancing protandry-based sorting, and lay desiccation-sensitive eggs, lowering accidental female release risk. The strain is compatible with size-based separation, improving sexing accuracy through the integration of natural and engineered dimorphisms. To our knowledge, this represents the first engineered sex-linked selectable trait in mosquitoes based on endogenous genes, establishing a foundation for scalable GSS development.</p>

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Mosquito sex separation using complementation of selectable traits and engineered neo-sex chromosomes

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年12月11日

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  • Doron S. Y. Zaada,
  • Or Toren,
  • Flavia Krsticevic,
  • Daniella A. Haber,
  • Denys Gildman,
  • Noam Galpaz,
  • Irina Häcker,
  • Marc F. Schetelig,
  • Eric Marois,
  • Yael Arien,
  • Philippos A. Papathanos

摘要

Effective sex separation remains a critical challenge for mosquito genetic control. Genetic sexing strains (GSS) address this by linking maleness with selectable traits, enabling efficient female removal. Here, we present a versatile platform for GSS development in the invasive Aedes albopictus mosquito that integrates CRISPR-engineered selectable phenotypes with sex conversion via nix, the male-determining factor. As a proof-of-concept, we disrupt the yellow pigmentation gene and restore its function in males using nix-containing transgenes, producing a stable strain with yellow females and dark males. Beyond serving as a vivid marker, yellow confers added advantages: GSS females pupate later than wild females, enhancing protandry-based sorting, and lay desiccation-sensitive eggs, lowering accidental female release risk. The strain is compatible with size-based separation, improving sexing accuracy through the integration of natural and engineered dimorphisms. To our knowledge, this represents the first engineered sex-linked selectable trait in mosquitoes based on endogenous genes, establishing a foundation for scalable GSS development.