Deletion of splicing factor Cdc5 in Toxoplasma disrupts transcriptome integrity, induces abortive bradyzoite formation, and prevents acute infection in mice

Toxoplasma gondii, an apicomplexan parasite, has over 75% of its genes containing introns; however, the role of RNA splicing in regulating gene expression remains unclear. Here, we demonstrate that the pre-mRNA splicing factor Cdc5 is part of a large spliceosomal complex essential for maintaining the transcriptome integrity in Toxoplasma. TgCdc5 depletion results in splicing inhibition with widespread changes in gene expression affecting several parasite processes, including the lytic cycle, DNA replication and repair, and protein folding and degradation. Consequently, non-cystogenic RH TgCdc5-depleted parasites begin spontaneously differentiating from tachyzoites to slow-growing bradyzoites, evidenced by the differential expression of key developmental regulators; however, these early-stage bradyzoites are unable to survive, likely due to a deficiency in functional proteins necessary for their growth and maintenance. Furthermore, consistent with our in vitro findings, we demonstrate that TgCdc5 is essential for parasite survival in mice, as its depletion provides complete protection against acute infection. Interestingly, this attenuated growth mutant resulting from TgCdc5 depletion elicits a robust immune response that fully protects mice from future infections and offers partial protection during pregnancy. Overall, this study highlights the indispensable role of the splicing factor Cdc5 in preserving transcriptional homeostasis in the intron-rich genome of Toxoplasma.