Conditional genetic manipulation systems in mosquitoes: Progress, Challenges, and perspectives

Mosquitoes are major vectors of severe infectious diseases, including malaria, dengue fever, and chikungunya, and continue to pose a substantial threat to global public health. A comprehensive understanding of mosquito-pathogen interactions, disease transmission mechanisms, and ecological adaptations is essential for developing innovative strategies to control mosquito-borne diseases. In recent years, genetic manipulation technologies have greatly advanced functional genomics research in mosquitoes; however, their limited spatiotemporal precision restricts the functional characterization of essential genes, key developmental regulators, and tissue-specific genes, often due to embryonic or early developmental lethality, systemic editing effects, and complex phenotypic consequences. As an important strategy for precision genetic regulation, Conditional Genetic Manipulation Systems (CGMSs) utilize tissue-specific, developmental stage-specific, or externally inducible regulatory elements to achieve controlled expression of functional genetic components (e.g., Cas9), thereby enabling spatiotemporally precise genetic modifications. These systems provide a new research paradigm for investigating mosquito basic biology and developing genetic control technologies. This review systematically summarizes the major types, regulatory mechanisms, and current advances of CGMSs in mosquitoes, with particular emphasis on different regulatory elements and their applications in functional gene analysis, studies of pathogen transmission mechanisms, and vector control strategies. Furthermore, we discuss future directions for expanding editing systems and developing genetic manipulation resources, aiming to provide insights into the construction of precise genetic manipulation systems in mosquitoes and other insects and the advancement of next-generation genetic control technologies.