The inducible gametocyte producer (iGP1) strain is well-suited to produce both immature and mature gametocytes for subsequent drug sensitivity profiling.

Advancing toward malaria eradication requires compounds that target both asexual blood-stage parasites to treat the disease as well as gametocytes to reduce transmission. However, current methods for gametocyte production and subsequent drug sensitivity testing, especially for immature gametocytes, lack standardization and are labor-intensive. Here, we describe a simplified and reproducible workflow for assessing gametocytocidal activity using the inducible gametocyte producer 1 (iGP1) line combined with the luciferase-based BacTiter-GLO viability detection kit. This approach yielded highly synchronous immature gametocytes enabling a robust drug sensitivity assay (Z´ = 0.65 ± 0.25). Testing of established and novel antimalarial compounds against immature and mature iGP1-gametocytes showed activity profiles comparable to previously published results, validating our approach. Of note, chlorotonil A and boromycin showed potent low-nanomolar activity against both immature and mature gametocytes, while the histone deacetylase inhibitors DS-089 and DS-118 displayed moderate (sub-) micromolar activity. In summary, this workflow provides a reliable and user-friendly platform for the generation of immature and mature gametocytes and subsequent drug sensitivity profiling with the potential to facilitate the discovery of novel gametocytocidal compounds and dual-active antimalarial treatments.