Single-cell RNA sequencing reveals a signature of sexual commitment in malaria parasites

Probing Plasmodium falciparum sexual commitment at the single-cell level

“…Comparative analysis of our single-cell data with those from Poran et al 15 and the bulk RNA-seq dataset validated the results by revealing a significant overlap in genes with commitment-associated expression profiles, despite differences in experimental setup and sequencing pipelines ( Table 2 and Supplementary File 5). Poran et al 15 also observed an enrichment of AP2-G binding motifs upstream of commitment-associated genes.…”

“…This hypothesis is also reinforced by the recent scRNA-seq study 15 , and by previous work using plaque assays and progeny analysis with gametocyte-specific antibodies 10 . To validate these observations directly, we investigated sexual commitment in reporter parasites.…”

“…Apart from ap2-g , the only commonly activated gene detected by both studies was the merozoite antigen msrp1 28, 29 . msrp1 has an AP2-G-binding site in its upstream sequence 15 , suggesting that it is activated by (and therefore downstream of) AP2-G. We observed significant overlap between our scRNA-seq and the population RNA-seq data 16 , as the same experimental set-up was used in these two studies. Shared hits include two kinases, a histone methyl transferase and two additional ApiAP2 transcription factors, further supporting the hypothesis that intracellular signaling events translate nutrient availability into epigenetic changes that ultimately regulate AP2-G.…”

“…Poran et al 15 also observed an enrichment of AP2-G binding motifs upstream of commitment-associated genes. We observe a similar enrichment of genes with upstream AP2-G binding sites within 2 kb of the translational start site (as defined in Campbell et al 24 ) within our commitment signature gene set (117 out of 125; p = 0.002, Fishers exact test, indicating that a significant proportion of the loci we detected may be downstream of AP2-G activation.…”

“…Two studies have identified AP2-G as a transcription factor that is activated during sexual commitment and required for gametocyte formation 13, 14 . Initial analysis of gene expression in individual sexually committed cells using conditional AP2-G knock-down provided a first transcriptional signature upon AP2-G activation 15 . We have recently demonstrated that availability of the host-derived phospholipid lysophosphatidylcholine (LysoPC) acts as an environmental sensor for the regulation of sexual commitment in P. falciparum 16 .…”