Mosquitoes are blood-feeding insects and serve as the most important vectors for spreading human diseases such as malaria, yellow fever, dengue fever, and filariasis. The continued use of synthetic insecticides has resulted in resistance in mosquitoes. Synthetic insecticides are toxic and affect the environment by contaminating soil, water, and air, and then natural products may be an alternative to synthetic insecticides because they are effective, biodegradable, eco-friendly, and safe to environment. Botanical origin may serve as suitable alternative biocontrol techniques in the future. The present study was carried out to establish the larvicidal potential of leaf extracts of Gmelina asiatica and synthesized silver nanoparticles using aqueous leaf extract against late third instar larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. Larvae were exposed to varying concentrations of plant extracts and synthesized AgNPs for 24 h. The results were recorded from UV-visible spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis support the biosynthesis and characterization of AgNPs. The maximum efficacy was observed in synthesized AgNPs against the larvae of An. stephensi (lethal dose (LC₅₀) = 22.44 μg/mL; LC₉₀ 40.65 μg/mL), Ae. aegypti (LC₅₀ = 25.77 μg/mL; LC₉₀ 45.98 μg/mL), and C. quinquefasciatus (LC₅₀ = 27.83 μg/mL; LC₉₀ 48.92 μg/mL), respectively. No mortality was observed in the control. This is the first report on mosquito larvicidal activity of plant-synthesized nanoparticles. Thus, the use of G. asiatica to synthesize silver nanoparticles is a rapid, eco-friendly, and a single-step approach and the AgNps formed can be potential mosquito larvicidal agents.