Genotyping Plasmodium berghei parasites

The PbPP1c–mCherry line was obtained by single homologous recombination as previously described [14 (link)]. Similarly, a C-terminal mCherry-tagged PbI2 was generated by single homologous recombination of a 1436 pb region of PbI2 without the stop codon (Pr3–Pr2, Supplementary Materials Table S3 and Figure S1A) inserted into the pL1886 vector and NdeI-linearized before transfection into the PbGFP ANKA strain.
Genotyping was performed on total DNA from parental and transfected parasites extracted from schizont pellets using the KAPA Express Extract Kit (KAPA BioSystem Inc, Wilmington, MA, USA). Transfected parasites were genotyped by diagnostic PCR using primers indicated in the Supplementary Materials Figure S1A and Table S3: Pr1–Pr2 for wild locus detection and Pr1–Pr4 for 3’ integration.