{"id":194565,"date":"2024-08-14T19:27:21","date_gmt":"2024-08-15T00:27:21","guid":{"rendered":"https:\/\/lifeboat.com\/blog\/2024\/08\/engineering-toxoplasma-gondii-secretion-systems-for-intracellular-delivery-of-multiple-large-therapeutic-proteins-to-neurons"},"modified":"2024-08-14T19:27:21","modified_gmt":"2024-08-15T00:27:21","slug":"engineering-toxoplasma-gondii-secretion-systems-for-intracellular-delivery-of-multiple-large-therapeutic-proteins-to-neurons","status":"publish","type":"post","link":"https:\/\/lifeboat.com\/blog\/2024\/08\/engineering-toxoplasma-gondii-secretion-systems-for-intracellular-delivery-of-multiple-large-therapeutic-proteins-to-neurons","title":{"rendered":"Engineering Toxoplasma gondii secretion systems for intracellular delivery of multiple large therapeutic proteins to neurons"},"content":{"rendered":"<p><a class=\"aligncenter blog-photo\" href=\"https:\/\/lifeboat.com\/blog.images\/engineering-toxoplasma-gondii-secretion-systems-for-intracellular-delivery-of-multiple-large-therapeutic-proteins-to-neurons.jpg\"><\/a><\/p>\n<p>Bracha <i>et al.<\/i><\/p>\n<hr>\n<p>\n<i><i>Toxoplasma<\/i> gondii<\/i> culture and maintenance.<\/p>\n<p>Type I RH and type II Pru and ME49 strain <i>T. gondii<\/i> were grown in HFF in high-glucose Dulbecco\u2019s modified Eagle\u2019s medium (DMEM) supplemented with 4 mM l-glutamine, 10% fetal bovine serum (FBS) and 1% penicillin\/streptomycin or 20 \u03bcg ml<sup>\u22121<\/sup> gentamicin antibiotics (\u2018complete DMEM\u2019) at 37 \u00b0C with 5% CO<sub>2<\/sub>. Cultures were monitored daily and <i>T. gondii<\/i> were passaged by transferring 1\u20133 drops (20\u2013100 \u03bcl) of the supernatant of a lysed dish (containing extracellular parasites) into a fresh dish with confluent HFF cells. Type I RH and type II Pru strains were validated by PCR\u2013restriction-fragment length polymorphism (primers described in Supplementary Table <a data-track=\"click\" data-track-label=\"link\" data-track-action=\"supplementary material anchor\" href=\"https:\/\/www.nature.com\/articles\/s41564-024-01750-6#MOESM2\">1<\/a>)<sup><a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 81\" title=\"Su, C., Zhang, X. & Dubey, J. P. Genotyping of Toxoplasma gondii by multilocus PCR-RFLP markers: a high resolution and simple method for identification of parasites. Int. J. Parasitol. 36841&ndash;848 (2006).\" href=\"https:\/\/www.nature.com\/articles\/s41564-024-01750-6#ref-CR81\" id=\"ref-link-section-d30852600e2689\">81<\/a><\/sup> or by passage into Cre Reporter cell lines to confirm Cre recombination as previously described<sup><a data-track=\"click\" data-track-action=\"reference anchor\" data-track-label=\"link\" data-test=\"citation-ref\" aria-label=\"Reference 16\" title=\"Koshy, A. A. et al. Toxoplasma secreting Cre recombinase for analysis of host&ndash;parasite interactions. Nat. Methods 7307&ndash;309 (2010).\" href=\"https:\/\/www.nature.com\/articles\/s41564-024-01750-6#ref-CR16\" id=\"ref-link-section-d30852600e2693\">16<\/a><\/sup>.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Bracha et al. Toxoplasma gondii culture and maintenance. Type I RH and type II Pru and ME49 strain T. gondii were grown in HFF in high-glucose Dulbecco\u2019s modified Eagle\u2019s medium (DMEM) supplemented with 4 mM l-glutamine, 10% fetal bovine serum (FBS) and 1% penicillin\/streptomycin or 20 \u03bcg ml\u22121 gentamicin antibiotics (\u2018complete DMEM\u2019) at 37 \u00b0C [\u2026]<\/p>\n","protected":false},"author":636,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[11,38,47],"tags":[],"class_list":["post-194565","post","type-post","status-publish","format-standard","hentry","category-biotech-medical","category-engineering","category-neuroscience"],"_links":{"self":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/194565","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/users\/636"}],"replies":[{"embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/comments?post=194565"}],"version-history":[{"count":0,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/posts\/194565\/revisions"}],"wp:attachment":[{"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/media?parent=194565"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/categories?post=194565"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/lifeboat.com\/blog\/wp-json\/wp\/v2\/tags?post=194565"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}