E-paper

Authors list: Zaada, DSY; Toren, O; Krsticevic, F; Haber, DA; Gildman, D; Galpaz, N; Häcker, I; Schetelig, MF; Marois, E; Arien, Y; Papathanos, PA

Publication year: 2025

Journal: BioRxiv

DOI Link: https://doi.org/10.1101/2025.03.30.646185

Publisher: Cold Spring Harbor Laboratory

Abstract: 

Effective and scalable sex separation remains a critical challenge for mosquito genetic control strategies. Genetic sexing strains (GSS) address this by genetically linking maleness with selectable traits, enabling efficient removal of females before release. Here, we describe a robust platform for the development of GSSs in the invasive Aedes albopictus mosquito by integrating a CRISPR-engineered selectable phenotype with sex conversion via nix, the male-determining factor. As a proof-of-concept, we disrupt the yellow gene to generate a vivid pigmentation marker, then rescue its function in males using nix-containing transgenes, creating a stable strain where all females are yellow and all engineered males are dark. The resulting GSS males are fertile, robust, and despite lacking the ancestral M locus, exhibit gene expression profiles closely resembling wild-type males. We benchmark sex separation based on pigmentation and discover that yellow mutant females exhibit slower larval development, enhancing protandry-based sorting. The GSS strain is compatible with existing size-based sex sorting systems, allowing for improved separation accuracy through the integration of natural and engineered sexually dimorphic traits. Additionally, we find that GSS females lay desiccation-sensitive eggs, reducing the risk of accidental female releases. Our approach is the first to engineer a sex-linked selectable trait by precisely targeting an endogenous gene and restoring its function in males, establishing a versatile platform for GSS development in Aedes mosquitoes.

Harvard Citation style: Zaada, D., Toren, O., Krsticevic, F., Haber, D., Gildman, D., Galpaz, N., et al. (2025) Mosquito Sex Separation using Complementation of Selectable Traits and Engineered Neo-Sex Chromosomes [Preprint]. BioRxiv, Article 2025.03.30.646185. https://doi.org/10.1101/2025.03.30.646185

APA Citation style: Zaada, D., Toren, O., Krsticevic, F., Haber, D., Gildman, D., Galpaz, N., Häcker, I., Schetelig, M., Marois, E., Arien, Y., & Papathanos, P. (2025). Mosquito Sex Separation using Complementation of Selectable Traits and Engineered Neo-Sex Chromosomes. BioRxiv. https://doi.org/10.1101/2025.03.30.646185