A novel chemogenomic screening platform for scalable antimalarial drug target identification

Abstract

SummaryLarge-scale chemical-genetic screening, or chemogenomics, can faciliate rapid and scalable drug target identification. To establish a chemogenomic screen for antimalarial drug target identification, we leveraged ∼600Plasmodium bergheiartificial chromosomes (PbACs) encoding potential drug targets to generate a systematic overexpression library. PbACs were engineered with DNA barcodes, enabling their quantification within mixed pools using next generation sequencing (barcode sequencing or BarSeq). Pooled transfection of PbACs into the highly genetically tractablePlasmodium knowlesidemonstrated efficient vector uptake and transcription of encodedP. bergheigenes. Parasite pools were exposed to antimalarial candidates, with pilot screens probing for known gene-compound associations identifying their targets with high sensitivity. Screening antimalarial inhibitors with unknown mechanisms of action successfully identifiedpi4kas the target for one novel compound, which was subsequently validated usingin vitroevolution inPlasmodium falciparumparasites. This sensitive and scalable chemogenomics platform therefore represents a valuable early-stage tool for antimalarial target identification.