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. 2020 Jul 9;5(13):e135589. doi: 10.1172/jci.insight.135589

Figure 3. P.falciparum mei2– parasites have no defects in blood stage and mosquito stage development.

Figure 3

(A) The schematic depicts the generation of the P. falciparum mei2– parasite using CRISPR/Cas9-mediated gene editing. Primers used to verify the gene deletion are indicated, and the sizes of the PCR products are shown in kilobases. Agarose gel electrophoresis shows the PCR products corresponding to the gene deletion P. falciparum mei2– clones F2 and F5. hDHFR, human dihydrofolate reductase. (B) The schematic (upper right) and the corresponding Southern blot for the P. falciparum NF54 and P. falciparum mei2– locus. The genomic DNA for P. falciparum NF54 and P. falciparum mei2– clones F2 and F5 was digested with XhoI, HindIII, and AflIII and probed with the 3′ Mei2 probe. (C) Asexual blood stage parasitemia is compared between P. falciparum NF54 (gray line) and P. falciparum mei2– clones F2 (red line) and F5 (blue line) over 3 replication cycles. Data represent mean ± SD. n = 3 biological replicates. Statistical analysis was carried out using 2-way ANOVA. ns, not significant. P > 0.05 is taken as ns. Graphs comparing (D) the counts for oocyst/midgut, (E) oocyst prevalence, and (F) counts for sporozoites/mosquito for P. falciparum NF54 (gray) and P. falciparum mei2– clones F2 (red) and F5 (blue). Data represent mean ± SD. n = 3 biological replicates. Statistical analysis was carried out using 1-way ANOVA. P > 0.05 is taken as ns.