Mutant/mutation
The mutant lacks expression of PY17X_1424200

Protein (function)
To identify membrane proteins critical in sensing XA or transducing XAinduced signal during gametogenesis, we identified 59 P. yoelii genes that are expressed in gametocytes and encode proteins with 1 to 22 predicted transmembrane domains (TMs) from the PlasmoDB database. We designed single guide RNA (sgRNA) to disrupt each of these genes using CRISPR/Cas9 methods16,17 and were able to successfully knockout (KO) 45 (76%) of the genes in the P. yoelii 17XNL strain, obtaining at least two cloned lines for each mutant . The remaining 14 genes (24%) were refractory to repeated deletion attempts using three independent sgRNA sequences, suggesting their essential roles for asexual blood-stage growth. The 45 gene deletion mutants proliferated asexually in mouse blood normally and were able to produce both male and female gametocytes although the gametocytemia level varied among these mutants.
Next we measured the gametogenesis of male gametocyte by counting exflagellation centers (ECs) formed in vitro after stimulation with 50 μMXAat 22 °C.Only one mutant (PY17X_1116300; GEP1) showed complete deficiency in EC formation and male gamete release and this mutant was analysed in greater detail.

Phenotype
Blood stage growth, gametocytemia and exflagellation was analysed.

Normal asexual blood stage growth/multiplication; normal production of gametocytes; normal exflagellation

Additional information

Other mutants
See other mutants generated in this study