Synthetic transcriptional control in the malaria parasite Plasmodium falciparum

Malaria is responsible for over half a million deaths each year. However, our understanding of malaria parasite biology is hampered by a lack of molecular tools, particularly at the level of transcriptional control. In light of this, we have created two orthogonal systems for inducible transcriptional repression in the malaria parasite Plasmodium falciparum using bacterial repressor proteins. We achieve 200- to 800-fold repression of expression, improving on previous attempts at transcriptional regulation by two orders of magnitude and outperforming gold standard translational/post-transcriptional regulation systems. We developed automated DNA design software to apply this tool to conditional regulation of native gene expression, validating essentiality and chemogenetic interactions with both two parasite lipid kinases and PfKelch13, which is associated with artemisinin resistance. These tools can advance our understanding and engineering of malaria functional genomics, drug mechanisms, and gene regulation.