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Previous work has demonstrated that\n                    <jats:italic>Toxoplasma gondii<\/jats:italic>\n                    , a parasite that naturally travels from the human gut to the central nervous system (CNS), can deliver proteins to host cells. Here we engineered\n                    <jats:italic>T. gondii<\/jats:italic>\n                    \u2019s endogenous secretion systems, the rhoptries and dense granules, to deliver multiple large (&gt;100\u2009kDa) therapeutic proteins into neurons via translational fusions to toxofilin and GRA16. We demonstrate delivery in cultured cells, brain organoids and in vivo, and probe protein activity using imaging, pull-down assays, scRNA-seq and fluorescent reporters. We demonstrate robust delivery after intraperitoneal administration in mice and characterize 3D distribution throughout the brain. As proof of concept, we demonstrate GRA16-mediated brain delivery of the MeCP2 protein, a putative therapeutic target for Rett syndrome. By characterizing the potential and current limitations of the system, we aim to guide future improvements that will be required for broader application.\n                  <\/jats:p>","DOI":"10.1038\/s41564-024-01750-6","type":"journal-article","created":{"date-parts":[[2024,7,29]],"date-time":"2024-07-29T12:04:07Z","timestamp":1722254647000},"page":"2051-2072","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Engineering Toxoplasma gondii secretion systems for intracellular delivery of multiple large therapeutic proteins to neurons"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5320-3352","authenticated-orcid":false,"given":"Shahar","family":"Bracha","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah 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