Development of CRISPR/Cas9-Based Genetic Strategies for Controlling Vector Populations (Aedes aegypti) to Mitigate Insecticide Resistance and Minimize Environmental Impact

Introduction: Vector-borne diseases, such as dengue, transmitted by Aedes aegypti, pose significant global health challenges, exacerbated by insecticide resistance and environmental concerns associated with conventional control methods. This study explores CRISPR/Cas9-based genetic strategies to suppress mosquito populations and block pathogen transmission while minimizing ecological impacts. Materials and Methods: Transgenic Aedes aegypti lines were generated using CRISPR/Cas9 to introduce sterility-inducing mutations in the doublesex (dsx) gene and pathogen-blocking mutations in the Niemann-Pick C1-like (NPC1) gene. Laboratory experiments assessed reproductive suppression and dengue virus (DENV-2) transmission, while mesocosm trials evaluated population dynamics and ecological effects. Insecticide resistance was tested via WHO bioassays, and mathematical modeling predicted long-term outcomes. Results: CRISPR/Cas9 achieved 78.3% and 85.1% mutation rates in dsx and NPC1 genes, respectively, with minimal off-target effects. Transgenic mosquitoes reduced egg production by 91.4% and DENV-2 transmission by 95.3%. Mesocosm trials showed 78.9% population suppression and no significant ecological disruptions. Insecticide susceptibility increased significantly in transgenic lines. Discussion: These findings demonstrate the efficacy and ecological safety of CRISPR-based vector control, offering a sustainable alternative to traditional methods. Future research should optimize gene drive efficiency and address regulatory challenges for field implementation.