{
  "abstract": "Transitioning from quantitative abundance measurements to functional activity-based assays represents a key opportunity in DNA repair research, particularly for enzymes like MUTYH that currently lack functional assay platforms. Here, we developed a FRET-based DNA switch for real-time monitoring of MUTYH glycosylase activity. The dumbbell-like architecture incorporates an 8-oxoG: A mismatch that leads to structure switching upon MUTYH-mediated adenine excision, resulting in activity-dependent fluorescence signal change. We demonstrate that the rationally designed GC-rich DNA nanoswitch can detect both E. coli MutY and human MUTYH with high specificity, enabling real-time activity monitoring directly in bacterial free cell extracts. In addition, the assay can be easily adapted for inhibitor screening, demonstrating its potential for drug screening applications. The activity-based assay was also implemented in HeLa cells and analyzed by confocal microscopy, confirming preferential localization of the MUTYH activity within the nuclear compartment. Overall, this simple, low-cost DNA-based sensor bridges the need for functional assays by directly monitoring MUTYH glycosylase activity in complex biological environments, offering a scalable platform for future screening applications.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Chemical Science and Technologies, University of Rome, Tor Vergata, Via della Ricerca Scientifica 1, Rome, Italy, 00133",
    "Department of Chemical and Biomolecular Engineering",
    "University of Melbourne",
    "Institute of Biosciences and BioResoures, National Research Council of Italy, Via Pietro Castellino, Naples, Italy"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 14,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Erica Belforte",
      "orcid": "0009-0009-0235-7971"
    },
    {
      "affs": [
        1,
        2
      ],
      "name": "Soraia Fernandes"
    },
    {
      "affs": [
        0
      ],
      "name": "Andrea Celeste Di Pede"
    },
    {
      "affs": [
        0
      ],
      "name": "Neda Bagheri"
    },
    {
      "affs": [
        0
      ],
      "name": "Laura Beltran Chamorro"
    },
    {
      "affs": [
        0
      ],
      "name": "Ankita Sinha"
    },
    {
      "affs": [
        0
      ],
      "name": "Raj Paul"
    },
    {
      "affs": [
        3
      ],
      "name": "Rosanna Mattossovich"
    },
    {
      "affs": [
        3
      ],
      "name": "Rosa Merlo"
    },
    {
      "affs": [
        0
      ],
      "name": "Giorgia Gambardella"
    },
    {
      "affs": [
        1,
        2
      ],
      "name": "Frank Caruso"
    },
    {
      "affs": [
        0,
        1,
        2
      ],
      "name": "Francesca Cavalieri"
    },
    {
      "affs": [
        3
      ],
      "name": "Anna Valenti"
    },
    {
      "affs": [
        0
      ],
      "name": "Alessandro Porchetta"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/frwz1c6njqe4m.bib",
  "cited_by": 0,
  "doi": "10.26434/chemrxiv.15010973/v1",
  "fulltext_url": null,
  "id": "frwz1c6njqe4m",
  "issns": [],
  "landing_url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010973/v1",
  "language": null,
  "licence": "CC-BY-NC",
  "licence_label": "CC BY-NC",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by-nc/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Advanced biosensing and bioanalysis techniques"
    ]
  ],
  "openalex": "W7223400945",
  "pdf_url": "https://chemrxiv.org/doi/pdf/10.26434/chemrxiv.15010973/v1",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-23",
  "publisher": "American Chemical Society (ACS)",
  "ris": "https://research.draconitic.com/paper/frwz1c6njqe4m.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:26Z",
      "native_id": "10.26434/chemrxiv.15010973",
      "source": "chemrxiv",
      "url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010973/v1"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Advanced biosensing and bioanalysis techniques",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Activity-based Detection of MUTYH using Engineered Dumbbell DNA Switches",
  "title_text": "Activity-based Detection of MUTYH using Engineered Dumbbell DNA Switches",
  "type": "preprint",
  "updated_at": "2026-10-30T12:17:26Z",
  "url": "https://research.draconitic.com/paper/frwz1c6njqe4m",
  "venue": "ChemRxiv",
  "versions": []
}