Functional validation of the Plasmodium falciparum K13 C580Y mutation in recently collected Ethiopian isolates

Abstract Recent genomic surveillance in Ethiopia identified the first detection of the Plasmodium falciparum kelch13 (K13) C580Y substitution in the Horn of Africa. To assess its functional impact, we introduced C580Y into two recently collected Ethiopian clinical isolates using CRISPR-Cas9 genome editing. Ring-stage survival assays demonstrated significantly elevated in vitro dihydroartemisinin (DHA) survival in edited parasites relative to isogenic controls, establishing that C580Y confers artemisinin tolerance in contemporary Ethiopian genetic backgrounds, providing one of the first causal assessments of C580Y in recent African parasite isolates.