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A process that is poorly understood. Through single-cell RNA profiling of\n                  <jats:italic>in<\/jats:italic>\n                  vivo-derived bradyzoites, we discovered that they are heterogeneous and not G1-arrested, as expected from\n                  <jats:italic>in vitro<\/jats:italic>\n                  studies. Instead, they progress through two cell cycles that branch from a single G1. While in G1b,\n                  <jats:italic>in vivo<\/jats:italic>\n                  -derived bradyzoites express cyst wall proteins predicted to replenish the wall that shields cysts. One cell cycle is common amongst tachyzoites and\n                  <jats:italic>in vitro-<\/jats:italic>\n                  and\n                  <jats:italic>in<\/jats:italic>\n                  vivo-derived bradyzoites; the other is unique to\n                  <jats:italic>in<\/jats:italic>\n                  vivo-derived bradyzoites (BCC). We demonstrated that AP2XI-6, expressed in G1\n                  <jats:italic>in vivo,<\/jats:italic>\n                  is dispensable for tachyzoite growth and cyst formation\n                  <jats:italic>in vitro<\/jats:italic>\n                  but necessary for encystation\n                  <jats:italic>in vivo.<\/jats:italic>\n                  We propose that AP2XI-6 is a driver of replication through the BCC that is required for cyst maturation and persistence.\n                <\/jats:p>","DOI":"10.1101\/2025.10.20.683543","type":"posted-content","created":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T04:10:14Z","timestamp":1761019814000},"source":"Crossref","is-referenced-by-count":1,"title":["Single-cell analysis of brain-derived\n                  <i>Toxoplasma<\/i>\n                  bradyzoites reveals a novel cell cycle regulated by AP2XI-6"],"prefix":"10.64898","author":[{"given":"Emma","family":"Franklin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0479-1300","authenticated-orcid":false,"given":"Argenis Arriojas","family":"Maldonado","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyra","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grace","family":"Hilliard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9983-1856","authenticated-orcid":false,"given":"Bruno Martorelli","family":"Di Genova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gary E","family":"Ward","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5108-8108","authenticated-orcid":false,"given":"Kourosh","family":"Zarringhalam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robyn","family":"Kent","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025102901524184000_2025.10.20.683543v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(04)16412-X"},{"key":"2025102901524184000_2025.10.20.683543v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/S1471-4922(02)02248-1"},{"key":"2025102901524184000_2025.10.20.683543v1.3","doi-asserted-by":"publisher","DOI":"10.1128\/EC.3.4.992-1003.2004"},{"key":"2025102901524184000_2025.10.20.683543v1.4","doi-asserted-by":"publisher","DOI":"10.1007\/s00436-010-1899-6"},{"key":"2025102901524184000_2025.10.20.683543v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.12.013"},{"issue":"5","key":"2025102901524184000_2025.10.20.683543v1.6","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1038\/s41564-023-01358-2","article-title":"A positive feedback loop controls Toxoplasma chronic differentiation","volume":"8","year":"2023","journal-title":"Nat Microbial"},{"issue":"9","key":"2025102901524184000_2025.10.20.683543v1.7","doi-asserted-by":"crossref","first-page":"e0064124","DOI":"10.1128\/mbio.00641-24","article-title":"Constitutive upregulation of transcription factors underlies permissive bradyzoite differentiation in a natural isolate of Toxoplasma gondii","volume":"15","year":"2024","journal-title":"mBio"},{"key":"2025102901524184000_2025.10.20.683543v1.8","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-40948-w"},{"key":"2025102901524184000_2025.10.20.683543v1.9","doi-asserted-by":"publisher","DOI":"10.2307\/3284713"},{"key":"2025102901524184000_2025.10.20.683543v1.10","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.1.501-507.2001"},{"key":"2025102901524184000_2025.10.20.683543v1.11","doi-asserted-by":"crossref","unstructured":"Zhang, Y.W. , et al., A Purification Method for Enrichment of the Toxoplasma gondii Cyst Wall. 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