{
  "abstract": "The antiviral innate immune response is a major species barrier that emerging viruses must overcome to infect and transmit efficiently in new hosts. In humans, the interferon-stimulated MX1 gene encodes the antiviral MX1 protein, which restricts zoonotic influenza A viruses (IAVs). Resistance to MX1 is considered a key feature of human-adapted IAVs and a potential determinant of zoonotic and pandemic risk. However, MX1-mediated restriction and viral escape have mostly been investigated in cell lines and animal models, which incompletely recapitulate virus-host interactions in the human airway epithelium, representing the primary site of IAV replication. Although primary human airway epithelial cell (hAEC) cultures, such as airway organoids and air-liquid interface (ALI) cultures, provide a physiologically relevant model, their use in studying MX1 restriction has been limited by challenges in achieving efficient and reliable genetic modification. Here, we used engineered virus-like particles (eVLPs) to deliver Cas9-sgRNA ribonucleoprotein complexes into primary hAECs and disrupt MX1 expression. This approach generated MX1-knockout airway organoids and organoid-derived hAEC ALI cultures with >90% genomic-level editing efficiency and near-complete loss of MX1 protein, while preserving epithelial differentiation at the air-liquid interface. MX1 knockout in hAEC ALI cultures had only a minor effect on the replication kinetics of the human-adapted H1N1pdm09 strain. By contrast, MX1 knockout markedly impaired interferon lambda-mediated restriction of the zoonotic bovine H5N1/Texas and avian H7N9/Anhui strains, resulting in viral titers up to 3-4 logs TCID50 higher than in MX1-competent cultures. These findings provide direct evidence that MX1 restricts zoonotic IAVs in primary human airway epithelium. Our study establishes a biologically relevant platform for assessing the sensitivity of emerging IAVs to a key antiviral host factor and thereby informs evaluations into their zoonotic and pandemic risk.",
  "abstract_shown": true,
  "affiliations": [
    "Institute for Infectious Diseases, University of Bern, Bern, Switzerland"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 15,
  "authors": [
    {
      "name": "K. Stevers"
    },
    {
      "name": "B. I. Oliveira Esteves"
    },
    {
      "name": "E. Aguiar Moreira"
    },
    {
      "name": "S. Jordi"
    },
    {
      "name": "L. Laloli"
    },
    {
      "name": "C. Moscardelli"
    },
    {
      "name": "M. F. Licheri"
    },
    {
      "name": "D. Wettstein"
    },
    {
      "name": "A. Mescoli"
    },
    {
      "name": "M. Mwanga"
    },
    {
      "name": "A. T. Baltensperger"
    },
    {
      "name": "L. Oberholster"
    },
    {
      "name": "P. V'kovski"
    },
    {
      "name": "G. Zimmer"
    },
    {
      "affs": [
        0
      ],
      "name": "R. Dijkman"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/46b7mdyyycam7.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.10.19.759965",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759965v1.full",
  "id": "46b7mdyyycam7",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759965v1",
  "language": null,
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "microbiology"
    ],
    [
      "openalex",
      "2713|Influenza Virus Research Studies"
    ]
  ],
  "openalex": "W7222931073",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759965v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-19",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/46b7mdyyycam7.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:28:38Z",
      "native_id": "10.64898/2026.10.19.759965",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.10.19.759965v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:microbiology",
      "subfield": "microbiology"
    }
  ],
  "title": "Genetic modification of primary airway epithelial cell cultures defines MX1 as a barrier for zoonotic influenza A viruses in the human respiratory epithelium",
  "title_text": "Genetic modification of primary airway epithelial cell cultures defines MX1 as a barrier for zoonotic influenza A viruses in the human respiratory epithelium",
  "type": "preprint",
  "updated_at": "2026-10-30T12:28:38Z",
  "url": "https://research.draconitic.com/paper/46b7mdyyycam7",
  "venue": "bioRxiv",
  "versions": []
}