{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T15:01:53Z","timestamp":1786978913046,"version":"build-2736575974"},"posted":{"date-parts":[[2026,8,13]]},"group-title":"Synthetic Biology","reference-count":63,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:00:00Z","timestamp":1786579200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Cancer Institute","award":["R01CA292860"],"award-info":[{"award-number":["R01CA292860"]}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["R35GM147478"],"award-info":[{"award-number":["R35GM147478"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["T32-AI153020"],"award-info":[{"award-number":["T32-AI153020"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,13]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Engineered bacteria offer a distinct modality for cancer therapy by exploiting the ability of certain species to colonize tumors and deliver therapeutic payloads. Improving their efficacy and safety requires control over bacterial activity after tumor colonization, yet few microbial chassis permit it.\n                  <jats:italic>Bifidobacterium longum<\/jats:italic>\n                  , a probiotic with intrinsic tumor-targeting and antitumor activity, is a promising chassis but lacks such control. Here, we develop a genetic control system that regulates\n                  <jats:italic>B. longum<\/jats:italic>\n                  activity within tumors, from gene expression to bacterial abundance. A human-isolate-derived replicon supports plasmid maintenance without antibiotic selection, and promoter and ribosome-binding-site libraries provide \u223c150-fold and \u223c48-fold expression ranges, respectively. Signal peptides enable secretion of structurally diverse therapeutic payloads and\n                  <jats:italic>B. longum<\/jats:italic>\n                  secreting CCL21 or an anti-PD-L1 nanobody reduces tumor growth relative to PBS controls. Anhydrotetracycline delivered in drinking water induces transgene expression in tumor-resident bacteria and reduces intratumoral bacterial load through CRISPRi targeting essential genes. Together, these results establish a tumor-homing probiotic as an externally controllable therapeutic chassis.\n                <\/jats:p>","DOI":"10.64898\/2026.08.12.744520","type":"posted-content","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T01:40:20Z","timestamp":1786671620000},"source":"Crossref","is-referenced-by-count":0,"title":["Programmable genetic control of tumor-colonizing\n                  <i>Bifidobacterium longum<\/i>\n                  for intratumoral therapeutic delivery and biocontainment"],"prefix":"10.64898","author":[{"given":"Jaehyun","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Microbiology, University of Chicago, Chicago, IL 60637, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua","family":"Glazier","sequence":"additional","affiliation":[{"name":"Pritzker School of Molecular Engineering, University of Chicago, Chicago, IL 60637, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ralph R.","family":"Weichselbaum","sequence":"additional","affiliation":[{"name":"Department of Radiation and Cellular Oncology, University of Chicago, Chicago, 60637, USA;"},{"name":"The Ludwig Center for Metastasis Research, University of Chicago, Chicago, 60637, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3083-2671","authenticated-orcid":false,"given":"Mark","family":"Mimee","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Chicago, Chicago, IL 60637, USA"},{"name":"Pritzker School of Molecular Engineering, University of Chicago, Chicago, IL 60637, USA"},{"name":"Duchoissois Family Institute, University of Chicago, Chicago, IL 60637, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026081707200517000_2026.08.12.744520v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc2934"},{"key":"2026081707200517000_2026.08.12.744520v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/s41568-018-0070-z"},{"key":"2026081707200517000_2026.08.12.744520v1.3","doi-asserted-by":"publisher","DOI":"10.1126\/science.add9667"},{"key":"2026081707200517000_2026.08.12.744520v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.addr.2025.115579"},{"key":"2026081707200517000_2026.08.12.744520v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-024-02418-6"},{"key":"2026081707200517000_2026.08.12.744520v1.6","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adc9436"},{"key":"2026081707200517000_2026.08.12.744520v1.7","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1007\/s00262-008-0555-9","article-title":"Salmonella typhimurium engineered to produce CCL21 inhibit tumor growth. 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