MYST-mediated epigenetic regulation determines the adaptive artemisinin-resistant in Plasmodium falciparum [ChIP-Seq]
To investigate the artemisinin resistance mechanism, we conducted a systematical evaluation of histone acetyltransferase expression in 45 cloned P. falciparum parasites and 30 wild-type field isolates. Remarkably, PfMYST, a member of the histone acetyltransferase MYST family, emerged as the sole candidate significantly associated with prolonged ring-survive of parasites. CHIP-seq analysis revealed PfMYST's pivotal role in mediating histone modifications, particularly in H4K5ac and H4K8ac, within the P. falciparum genome. Through single-cell RNA sequence and conditional knockdown approaches, we identified and functionally validated PfMYST-targeted genes contributing to Plasmodium's adaptive artemisinin resistance. Overall design: We generated transgenic parasite lines tagged with HA and GFP (PfMYST-HA_GFP) in 3D7 stain using the CRISPR-Cas9 technique. To explore changes in histone acetylation upon PfMYST knockdown, we conducted a series of chromatin immunoprecipitation and high-throughput sequencing (ChIP-seq) assays targeting PfMYST, H4K5ac, H4K8ac, H4K12ac, and H4K16ac in both ring-stage and schizont-stage of PfMYST-HA_GFP and PfMYST-Ribo (PfMYST-off and wild type clones) respectively.