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However, most of the known effector proteins are specific to <jats:italic>Toxoplasma<\/jats:italic> genotypes or hosts. To identify virulence factors that function across different parasite isolates and mouse strains that differ in susceptibility to infection, we performed systematic pooled in vivo CRISPR-Cas9 screens targeting the <jats:italic>Toxoplasma<\/jats:italic> secretome. We identified several proteins required for infection across parasite strains and mouse species, of which the dense granule protein 12 (GRA12) emerged as the most important effector protein during acute infection. GRA12 deletion in IFN\u03b3-activated macrophages results in collapsed parasitophorous vacuoles and increased host cell necrosis, which is partially rescued by inhibiting early parasite egress. GRA12 orthologues from related coccidian parasites, including <jats:italic>Neospora caninum<\/jats:italic> and <jats:italic>Hammondia hammondi<\/jats:italic>, complement Tg\u0394GRA12 in vitro, suggesting a common mechanism of protection from immune clearance by their hosts.<\/jats:p>","DOI":"10.1038\/s41467-025-58876-2","type":"journal-article","created":{"date-parts":[[2025,4,16]],"date-time":"2025-04-16T09:03:13Z","timestamp":1744794193000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["GRA12 is a common virulence factor across Toxoplasma gondii strains and mouse subspecies"],"prefix":"10.1038","volume":"16","author":[{"given":"Francesca","family":"Torelli","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4605-291X","authenticated-orcid":false,"given":"Simon","family":"Butterworth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eloise","family":"Lockyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-9534-0248","authenticated-orcid":false,"given":"Ana N.","family":"Matias","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franziska","family":"Hildebrandt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8462-2741","authenticated-orcid":false,"given":"Ok-Ryul","family":"Song","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Pearson-Farr","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7883-6743","authenticated-orcid":false,"given":"Moritz","family":"Treeck","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,4,16]]},"reference":[{"key":"58876_CR1","doi-asserted-by":"publisher","first-page":"1385","DOI":"10.1016\/j.ijpara.2009.04.003","volume":"39","author":"G Pappas","year":"2009","unstructured":"Pappas, G., Roussos, N. & Falagas, M. 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