Mapping Plasmodium transitions and interactions in the Anopheles female

Gene expression during the late ookinete and oocyst stages of Plasmodium falciparum (human malaria parasite) development has not been previously described in detail. We used single-cell RNA-seq to map gene expression changes in both parasite and mosquito cells and their interactions at multiple timepoints after an infectious blood meal, and in control (dsGFP-injected) and EcR-depleted (dsEcR-injected) mosquitoes. Infected midguts were dissected from 80 dsRNA-injected female mosquitoes (dsGFP or dsEcR) and cellularlized by enzymatic digestion, purified and analyzed by single-cell RNA-seq (10X Genomics). Timepoints were 36 h, 2 d, 4 d, and 7 d post infectious blood meal. On-day old mosquitoes were injected with dsRNA 3 d prior to blood feeding.