Development of a bsd‐blasticidin selection system in Plasmodium berghei

Plasmodium berghei is used as a rodent model for the study of malaria. However, multiple genetic manipulations are restricted by the paucity of selectable markers. The bsd‐blasticidin selection system is widely used for eukaryotic cells; however, it could not previously be used for P. berghei due to toxicity to the rodent host. Here, we report the application of this selection system in P. berghei using an in vitro selection method. The desired bsd‐integrated mutants are enriched by more than 90% within 2 weeks when using this system. Furthermore, the bsd marker can be used sequentially with established pyrimethamine‐ and puromycin‐resistant markers. This system allows deeper understanding of malaria parasite biology through extensive genetic manipulation of P. berghei.