A single-cell atlas of Toxoplasma sexual development in the feline intestinal tract

Toxoplasma gondii undergoes sexual development exclusively in the feline intestine, a process critical for genetic diversity and population expansion. Recent studies have identified genes critical in suppressing presexual development and metabolic differences in felines that may promote sexual development, but to date, the gene regulatory networks driving development in the cat are unknown. To investigate this, we performed single-cell transcriptomics on parasites isolated from cat intestines, using fluorescent reporter strains and flow cytometry. From 15,068 cells across 2 experiments, we identified rare populations, including cells that bear all of the hallmarks of gametes. Candidate genes emerging from this study were tested via CRISPR–Cas9 perturb-seq, identifying AP2X6 as a regulator of macrogametocyte development. Our single-cell data extend what is known about gene expression changes throughout sexual development and should be useful to those in the field working towards inducing gametogenesis, mating and oocyst production in vitro. Single-cell RNA sequencing is used to study the sexual development of Toxoplasma gondii in cats, generating a transcriptional atlas and showing that the female-specific gene AP2X6 functions as a regulator of oocyst formation. Frenkel, J. K. & Dubey, J. P. Toxoplasmosis and its prevention in cats and man. J. Infect. Dis. 126, 664–673 (1972). Dubey, J. P. Oocyst shedding by cats fed isolated bradyzoites and comparison of infectivity of bradyzoites of the VEG strain Toxoplas... [9643 chars]