Figure 3: Schematic of the ZFN-based pfmdr1 editing strategy. | Nature Communications

Figure 3: Schematic of the ZFN-based pfmdr1 editing strategy.

From: Globally prevalent PfMDR1 mutations modulate Plasmodium falciparum susceptibility to artemisinin-based combination therapies

Figure 3

The pmdr1 plasmid expresses the 2A-linked pfmdr1-specific ZFN pair from the calmodulin promoter and the human dhfr selectable marker. These ZFNs create a double-stranded break that can be repaired using the plasmid-borne homologous donor sequence that extends 0.6 kb upstream and 1.8 kb downstream of the target site (yellow thunderbolt). The four pmdr1 transfection plasmids carry three silent mutations at the binding sites to prevent ZFNs cleaving the plasmids or the edited sequence. These plasmids encode the four haplotypes at residues 86 and 184. Shown is the example of transfecting a N86/Y184 parasite with the pmdr1NY plasmid (Table 2). Chromatograms show sequence analysis of genomic and recombinant DNA samples. Mutations at the ZFN binding site (red arrows) and the N86Y and Y184F change of codons are indicated.

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