{
  "abstract": "Endosomal escape limits RNA delivery by lipid nanoparticles (LNPs). If escape requires membrane rupture, productive delivery would be intrinsically coupled to membrane damage and its potential toxicity. Here, we establish a cell-free system based on giant unilamellar vesicles (GUVs) with an early endosome-like lipid composition to dissect the mechanism of siRNA transfer and determine whether it can occur while membrane barrier function is preserved. Acidification promoted LNP-membrane association, lipid redistribution and siRNA translocation into the receiving lumen without detectable dissipation of transmembrane pH gradient. Complementary cryo-electron tomography revealed local interfacial lipid reorganisation without detectable membrane rupture. Fluorescence correlation spectroscopy revealed heterogeneous diffusion of translocated siRNA, consistent with free and lipid-associated species. Pre-formed RNA-lipid complexes exhibited pH-dependent dissociation and heterogeneous diffusion, supporting lipid association as a possible origin of slower luminal diffusion. Serum or apolipoprotein-E preincubation reduced siRNA translocation into the GUV lumen. Together, these findings support a model where siRNA-lipid association facilitates membrane crossing without sustained loss of barrier function. This platform provides a means to dissect LNP-mediated RNA delivery and develop formulations with improved efficacy while limiting membrane damage.",
  "abstract_shown": true,
  "affiliations": [
    "Human Technopole"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "name": "A. Spadea"
    },
    {
      "name": "A. Singh"
    },
    {
      "name": "L. Redlingshoefer"
    },
    {
      "name": "S. Sorrentino"
    },
    {
      "name": "Y. Kalaidzidis"
    },
    {
      "affs": [
        0
      ],
      "name": "M. Zerial"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/0rdrcqnnj7xga.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.11.02.763664",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.11.02.763664v1.full",
  "id": "0rdrcqnnj7xga",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.11.02.763664v1",
  "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",
      "cell biology"
    ]
  ],
  "openalex": null,
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.11.02.763664v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-11-02",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/0rdrcqnnj7xga.ris",
  "sources": [
    {
      "first_seen": "2026-11-04T00:21:24Z",
      "native_id": "10.64898/2026.11.02.763664",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.11.02.763664v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:cell biology",
      "subfield": "cell-biology"
    }
  ],
  "title": "Reconstitution of lipid nanoparticle-mediated siRNA transfer across endosomal membrane mimics",
  "title_text": "Reconstitution of lipid nanoparticle-mediated siRNA transfer across endosomal membrane mimics",
  "type": "preprint",
  "updated_at": "2026-11-05T00:20:44Z",
  "url": "https://research.draconitic.com/paper/0rdrcqnnj7xga",
  "venue": "bioRxiv",
  "versions": []
}