Advancing mosquito control: Integrating SEPARATOR and CRISPR-based Strategies for Effective Biocontrol of Aedes aegypti

Female mosquitoes, as the primary blood-feeders, are responsible for transmitting dangerous human pathogens. This underscores the critical necessity for effective control strategies targeting these blood-feeding vectors. Previously, we introduced SEPARATOR (Sexing Element Produced by Alternative RNA-Splicing of A Transgenic Observable Reporter), a novel method designed to exploit sex-specific alternative splicing of a benign reporter gene, ensuring exclusive expression in males. Additionally, we developed the CRISPR-based precision-guided sterile insect technique (pgSIT) for Aedes aegypti, which disrupts genes essential for viability and fertility during development to genetically generate sex-sorted and viable sterilized males. We demonstrated that released pgSIT males can effectively compete with, suppress, and eliminate caged mosquito populations. In this study, we demonstrate the versatility of SEPARATOR when integrated with CRISPR-based mosquito control strategies. This integration promises streamlined production of male mosquitoes for release programs, crucial for genetic biocontrol interventions. The scalability and adaptability of SEPARATOR across species underscore its significance in mosquito population management. Our findings lay the groundwork for developing a refined sterile insect technique tailored for efficient mosquito control in Aedes aegypti populations, thereby advancing strategies to combat mosquito-borne diseases.