Figure 2.

Generation of 3D7^cas9. (A) The 3D7^pcen_cas9 parasite that maintained the pCen_cas9 plasmid was co-transfected with the Cas9 expression cassette and the psgRNA_kahrp_cen plasmid using their fully mature schizonts. The expression cassette was integrated into the kahrp locus. (B) Genotyping PCR of 3D7^cas9 parasites was performed using the p1 and p2 primers. Full length gel image is included in Fig. S4A. (C) Expression of Cas9 nuclease was confirmed by Western blot analysis using the anti-FLAG antibody. Histone H3 was used as an internal control. Full length blots are included in Fig. S4B. (D) The growth of 3D7^cas9 parasites (blue, circle) during asexual development in RBCs was comparable to that of wild-type parasites (black, box). Positive and negative errors were calculated from the standard error of the mean from biological triplicates. Distributions for each day were compared using the unpaired t-test (not significant).