Recent advances in identifying and characterizing secretory proteins of Toxoplasma gondii by CRISPR-based screening.

The apicomplexan parasite, Toxoplasma gondii, develops unique secretory organelles, such as micronemes, rhoptries, and dense granules, which do not exist in other well-studied eukaryotic organisms. These secretory organelles are key features of apicomplexan parasites and discharge various proteins that are essential for invasion, replication, egress, host-parasite interactions, and virulence. Many studies have therefore focused on identifying and characterizing the proteins secreted by T. gondii that play essential roles in pathology and that can be targeted for therapeutics and vaccine development. The recent development of functional genetic screens based on CRISPR/Cas9 technology has revolutionized this field and has enabled the identification of genes that contribute to parasite fitness in vitro and in vivo. Consequently, characterization of genes identified by unbiased CRISPR screens has revealed novel aspects of apicomplexan biology. In this review, we describe the development of CRIPSR-based screening technology for T. gondii, and recent advances in our understanding of secretory proteins identified and characterized by CRISPR-based screening.