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. 2021 May 19;6(3):e01093-20. doi: 10.1128/mSphere.01093-20

FIG 1.

FIG 1

Screening of Plasmodium falciparum nonessential kinases during sexual commitment and development. (A) List of nonessential kinases characterized during sexual development in this study (PF3D7_0107600, eIK2; PF3D7_0525900, NEK2; PF3D7_0615500, CRK5; PF3D7_0623800, TKL4; PF3D7_0715300, calcium/calmodulin-dependent protein kinase, putative; PF3D7_1121300, TKL2; PF3D7_1201600, NEK3; PF3D7_1423600, calcium-dependent protein kinase, putative; PF3D7_1450000, serine/threonine protein kinase, putative) (14). (B) Comparison of the percentage of gametocytemia between the 3D7 WT line and the PfTKL2 kinase KO clones of the same transfection (clones B10 and B12) generated by single crossover integration (14). Each column represents the mean of triplicate microscope counts, each of at least 500 cells, analyzed using paired t test, ± standard deviation (*, P < 0.05; 3D7 versus TKL2 KO clones, P = 0.0377). (C) Schematic of the CRISPR/Cas9 strategy used to generate a TKL2 conditional knockout (KO) line (3D7/TKL2:loxPint) as well as the primers used to confirm successful gene editing (Table S2). The pMK-RQ-tkl2-loxPint donor plasmid contains a recodonized version of the glycine loop in the kinase domains of tkl2 (rc. Gly oop) flanked by two loxPints and homology regions for homology-directed repair. (D) Sexual conversion rates in 3D7/TKL2:loxPint parasites treated with DMSO (control) or rapamycin (KO). Each column represents the mean of triplicate microscope counts, each of at least 500 cells, analyzed using paired t test ± SD, (ns, P ≥ 0.05; 3D7/TKL2:loxPint treated with DMSO versus rapamycin, P = 0.4017).