Single-cell transcriptomic profiling of Toxoplasma gondii eIF5A K49 mutants during alkaline stress-induced differentiation

Using the ME49 strain as the genetic background, we performed 10x Genomics single-cell RNA sequencing on wild-type parasites, an eIF5A-K49Q acetylation-mimic mutant, and an eIF5A-K49R non-acetylatable charge-conservative mutant.

Each parasite line was cultured under standard conditions or exposed to alkaline medium (pH 8.2 for 4 days) to induce tachyzoite-to-bradyzoite differentiation in vitro. For each genotype-by-condition group, three independently cultured biological replicates were pooled before single-cell capture to generate one scRNA-seq library. Six pooled libraries were generated in total.