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Two members of 6-cysteine (6-cys) protein family, P48/45 and P230, are important for gamete fertility in rodent and human malaria parasites and are leading transmission blocking vaccine antigens. Rodent and human parasites encode a paralog of P230, called P230p. While P230 is expressed in male and female parasites, P230p is expressed only in male gametocytes and gametes. In rodent malaria parasites this protein is dispensable throughout the complete life-cycle; however, its function in  P .  falciparum  is unknown. Using CRISPR/Cas9 methodology we disrupted the gene encoding  Pfp230p  resulting in  P .  falciparum  mutants ( Pf Δ p230p ) lacking P230p expression. The  Pf Δ p230p  mutants produced normal numbers of male and female gametocytes, which retained expression of P48/45 and P230. Upon activation male  PfΔp230p  gametocytes undergo exflagellation and form male gametes. However, male gametes are unable to attach to red blood cells resulting in the absence of characteristic exflagellation centres  in vitro . In the absence of P230p, zygote formation as well as oocyst and sporozoite development were strongly reduced (>98%) in mosquitoes. These observations demonstrate that P230p, like P230 and P48/45, has a vital role in  P .  falciparum  male fertility and zygote formation and warrants further investigation as a potential transmission blocking vaccine candidate.

The Plasmodium falciparum male gametocyte protein P230p, a paralog of P230, is vital for ookinete formation and mosquito transmission