Generation and characterization of the NF54::DiCre line for conditional gene deletions across the P. falciparum life cycle. (a) Schematics of the targeting (pBSPs47DiCre) plasmid, the expected recombination product, and representation of the primer pairs used to test integration. (b) PCR analysis of DiCre cassette integration into the Pfs47 locus in P. falciparum NF54. The sequences of the primers used (e.g., primer #151) are shown in Table S1 in the supplemental material. (c) Illustration of the P. falciparum life cycle and expected timeline of the different developmental stages addressed in this paper. (d) Percentage of A. stephensi mosquitoes infected with NF54 and NF54::DiCre after a blood meal, where each data point corresponds to the value from an independent experiment. (e) Number of NF54 and NF54::DiCre oocysts per mosquito gut. Each data point corresponds to the value from an independent experiment. (f) Comparison of hepatocyte infection rates between NF54 and NF54::DiCre parasites of at least four independent experiments, each with two technical replicates. All P values were calculated by the Mann-Whitney t test.