Unveiling Antimalarial Drug Resistance: The Role of CRISPR‐Cas Technologies in Malaria Research

Background Malaria, caused by Plasmodium parasites, remains a global health burden, with drug resistance threatening control efforts. CRISPR‐Cas technologies, particularly CRISPR‐Cas9, have transformed malaria research by enabling precise genetic manipulations to study antimalarial resistance mechanisms, develop novel therapeutics and enhance diagnostics. Methods A systematic literature search (2015–2025) across Web of Science, Scopus and PubMed identified 39 peer‐reviewed studies utilising CRISPR‐Cas to investigate antimalarial drug resistance in Plasmodium species . Data were extracted on study design, genetic manipulations, target genes, resistance mechanisms and outcomes. Results CRISPR‐Cas9 was the dominant tool (84.6%), primarily used for point mutations ( PfK13 C580Y, PfCRT K76T), gene knockouts and integrations in P. falciparum (87.2%) and P. berghei (12.8%). Key resistance mechanisms included drug efflux ( PfCRT , PfMDR1 ), reduced binding affinity ( PfDHODH , DHFR ), metabolic compensation ( PfDHODH CNV) and impaired haemoglobin trafficking ( PfK13 , UBP‐1). Novel mediators like PfCARL and PfExo were identified, alongside compensatory mutations mitigating fitness costs. High‐throughput screening (BarSeq) and diagnostics (SHERLOCK, CRISPR‐Cas 12a) enhanced resistance surveillance. Challenges included low editing efficiency in nonasexual stages, limited P. vivax tools, off‐target risks and scalability constraints. Conclusion CRISPR‐Cas technologies have elucidated critical resistance pathways and supported innovative diagnostics, but challenges persist. Future advancements in base editing, omics integration and in vivo models will expand CRISPR’s utility in nonfalciparum species and field isolates, informing novel antimalarials and surveillance strategies to combat drug‐resistant malaria.