{
  "abstract": "Engineered bacteria offer programmable cancer immunotherapies, but engineering tractability does not always align with therapeutic suitability. The attenuated intracellular vaccine strain Brucella melitensis {Delta}vjbR (Bm{Delta}vjbR) shows promise as an immunotherapy chassis, yet its translation requires reliable control and containment systems that remain difficult to engineer with the limited genetic tools available. Here, we establish a genetic domestication workflow that connects genetic part generation and quantitative evaluation with model-guided circuit design. Through mutagenesis and screening, we generated and characterized genetic part libraries, including promoters spanning two orders of magnitude in activity, a panel of ribosome-binding sites, and highly functional terminators. We also evaluated fluorescent reporters, plasmid compatibility, and three inducible regulatory systems. These host-specific measurements informed mathematical modeling to guide the selection of a doxycycline-inducible Cas9/gRNA kill-switch architecture before construction. Experimental validation demonstrated inducible control both in vitro and in vivo: doxycycline reduced viable Bm{Delta}vjbR below the assay detection limit in vitro and decreased bacterial burden in mouse liver by approximately 91%. To our knowledge, this work establishes the first systematic genetic domestication of Bm{Delta}vjbR and the first containment circuit in this strain. The resulting workflow provides an extensible framework for domesticating and engineering other non-model bacteria.",
  "abstract_shown": true,
  "affiliations": [
    "Texas A&M University"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "name": "Y. Jeong"
    },
    {
      "name": "H. R. Namboothiri"
    },
    {
      "name": "O. Femi-Olatunji"
    },
    {
      "name": "P. de Figueiredo"
    },
    {
      "name": "C. Agca"
    },
    {
      "affs": [
        0
      ],
      "name": "C. Y. Hu"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/2qwk5z564cjcq.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.10.18.759933",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.10.18.759933v1.full",
  "id": "2qwk5z564cjcq",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.10.18.759933v1",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "synthetic biology"
    ],
    [
      "openalex",
      "1305|Cancer Research and Treatments"
    ]
  ],
  "openalex": "W7223064285",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.10.18.759933v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-18",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/2qwk5z564cjcq.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:28:42Z",
      "native_id": "10.64898/2026.10.18.759933",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.10.18.759933v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:synthetic biology",
      "subfield": "systems-synthetic"
    }
  ],
  "title": "Integrating Genetic Domestication and Model-Guided Kill-Switch Engineering in a Non-Model Bacterial Therapeutic",
  "title_text": "Integrating Genetic Domestication and Model-Guided Kill-Switch Engineering in a Non-Model Bacterial Therapeutic",
  "type": "preprint",
  "updated_at": "2026-10-30T12:28:42Z",
  "url": "https://research.draconitic.com/paper/2qwk5z564cjcq",
  "venue": "bioRxiv",
  "versions": []
}