{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:02:00Z","timestamp":1785427320165,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,9]]},"group-title":"Microbiology","reference-count":67,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T00:00:00Z","timestamp":1783555200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004359","name":"Swedish Research Council","doi-asserted-by":"crossref","award":["2021-06749"],"award-info":[{"award-number":["2021-06749"]}],"id":[{"id":"10.13039\/501100004359","id-type":"DOI","asserted-by":"crossref"}]},{"award":["AI007413"],"award-info":[{"award-number":["AI007413"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["OD034245"],"award-info":[{"award-number":["OD034245"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["AI180278"],"award-info":[{"award-number":["AI180278"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["GM151129"],"award-info":[{"award-number":["GM151129"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Intracellular pathogens remodel host cells by redirecting cellular machinery to the host-pathogen interface. The protozoan parasite\n                  <jats:italic>Toxoplasma gondii<\/jats:italic>\n                  co-opts host ESCRT proteins at the parasitophorous vacuole membrane (PVM), where they support budding of host-derived vesicles into the vacuole. Yet the parasite effectors that direct this process remain largely unknown. Here we developed spaCR (\n                  <jats:bold>s<\/jats:bold>\n                  patial\n                  <jats:bold>p<\/jats:bold>\n                  henotype\n                  <jats:bold>a<\/jats:bold>\n                  nalysis of\n                  <jats:bold>CR<\/jats:bold>\n                  ISPR-Cas9 screens), a pooled image-based screening framework that combines deep-learning classification of single-cell spatial phenotypes with well-level barcode genotyping and regression-based deconvolution of gene effects. Screening\n                  <jats:italic>T. gondii<\/jats:italic>\n                  secretory proteins recovered the known TSG101 recruiter GRA14 and identified an uncharacterized effector, ESCRT-association factor 1 (EAF1), that promoted TSG101 recruitment to the PVM. Targeted deletion confirmed its role across Type I and II lineages, and IP-MS recovered ESCRT-I and ESCRT-III components, including TSG101. Together, these findings identify EAF1 as an ESCRT-associated parasite effector and establish spaCR as a general framework for linking CRISPR perturbations to spatial phenotypes.\n                <\/jats:p>","DOI":"10.64898\/2026.07.08.737057","type":"posted-content","created":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T14:05:17Z","timestamp":1783605917000},"source":"Crossref","is-referenced-by-count":0,"title":["A pooled image-based CRISPR screen identifies EAF1 as a\n                  <i>T. gondii<\/i>\n                  modulator of ESCRT subversion"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3388-061X","authenticated-orcid":false,"given":"Einar B.","family":"Olafsson","sequence":"first","affiliation":[{"name":"Department of Cell and Molecular Biology, Microbiology and Immunology, Uppsala University, Uppsala, Sweden"},{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2392-5837","authenticated-orcid":false,"given":"Christophe-Sebastien","family":"Arnold","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9211-7406","authenticated-orcid":false,"given":"Jacob A.","family":"Kellermeier","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-6834-8343","authenticated-orcid":false,"given":"Patrick A.","family":"Rimple","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8275-2091","authenticated-orcid":false,"given":"Hargobinder","family":"Kaur","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9451-0041","authenticated-orcid":false,"given":"Yifan","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9244-5888","authenticated-orcid":false,"given":"Jonathan Z.","family":"Sexton","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Gastroenterology and Hepatology, Michigan Medicine at the University of Michigan, Ann Arbor, MI, 48109, USA"},{"name":"Gilbert S. Omenn Department of Computational Medicine & Bioinformatics, University of Michigan, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7392-1746","authenticated-orcid":false,"given":"Staffan","family":"Sv\u00e4rd","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Biology, Microbiology and Immunology, Uppsala University, Uppsala, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6859-8895","authenticated-orcid":false,"given":"Vern B.","family":"Carruthers","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3128-5331","authenticated-orcid":false,"given":"Matthew J.","family":"O\u2019Meara","sequence":"additional","affiliation":[{"name":"Gilbert S. 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