Comparative RNA-BisSeq analysis on the schizont stages of 17XNL wild-type (WT) and Pynsun2 knock-out parasites demonstrated a marked decrease in the density of m5C modifications in mRNA transcripts. By RNA-BisSeq, 7409 m5C sites were detected in 527 mRNAs of the WT parasites whereas this number was reduced to 1845 m5C sites in 265 mRNAs of the knock-outs

Experiments with the knock-out and WT lines demonstrated that disruptions of Pynsun2 resulted in dramatically reduced gametocyte production. The disruption of Pynsun2 affected the production of male and female gametocytes similarly while it showed no detectable effect on the propagation of asexual blood-stage parasites.

Consistent with the effect of gene disruption on gametocyte production, in vitro ookinete conversion rates to mature forms were significantly reduced in the knock-outs relative to those of the WT parasites. Also, the numbers of midgut oocysts at day 7 and salivary gland sporozoites at day 14 were markedly decreased in Pynsun2 knock-out parasites, suggesting that Pynsun2 knock-out effects may impair the development of parasite stages in the mosquito

Consistent with the effect of gene disruption on gametocyte production, in vitro ookinete conversion rates to mature forms were significantly reduced in the knock-outs relative to those of the WT parasites. Also, the numbers of midgut oocysts at day 7 and salivary gland sporozoites at day 14 were markedly decreased in Pynsun2 knock-out parasites, suggesting that Pynsun2 knock-out effects may impair the development of parasite stages in the mosquito

Consistent with the effect of gene disruption on gametocyte production, in vitro ookinete conversion rates to mature forms were significantly reduced in the knock-outs relative to those of the WT parasites. Also, the numbers of midgut oocysts at day 7 and salivary gland sporozoites at day 14 were markedly decreased in Pynsun2 knock-out parasites, suggesting that Pynsun2 knock-out effects may impair the development of parasite stages in the mosquito