{
  "abstract": "The dCas9 system has rapidly been developed into many tools to explore different aspects of the human genome; the high binding specificity, coupled with the inactive nuclease enzyme, allows for precise recruitment of molecules to specific sequences of DNA. We sought to exploit these capabilities to create a tool for assessing the real-time proximity of two DNA sequences within a biological system. By incorporating aptamers into gRNAs, dCas9 molecules can be used to recruit the β9 or β10 strands of split-NanoLuc® to specific DNA sequences and quantify the proximity of those sequences based on their ability to complex with the luciferase fragment (Δ11S) and produce luminescence. While many tools exist to detect a single DNA sequence, this system is uniquely capable of assessing how two DNA sequences interact with each other. As expected, we found that the interaction of two dCas9 molecules was affected by their linear distance from each other on dsDNA. Surprisingly, we found that their interaction was also strongly influenced by rotational orientation, even for sequences that are close together in linear space. This finding indicates that dCas9 rotational alignment is an important consideration for designing dCas9 systems that target multiple DNA sequences simultaneously. Beyond the findings presented herein, we believe this DNA proximity detection tool has the potential to be adapted for applications involving the proximity and orientation of two DNA sequences.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Cell & Developmental Biology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States of America.",
    "Department of Molecular Pharmaceutics, University of Utah, Salt Lake City, Utah, United States of America.",
    "Department of Medicinal Chemistry; Department of Biomedical Engineering, University of Utah, Salt Lake City, Utah, United States of America.",
    "Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, Utah, United States of America."
  ],
  "article_type": "Research Article",
  "arxiv": null,
  "author_total": 4,
  "authors": [
    {
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        0
      ],
      "name": "Saba Parvez"
    },
    {
      "affs": [
        1
      ],
      "name": "Morgan C Marsh"
    },
    {
      "affs": [
        1,
        2
      ],
      "name": "Shawn C Owen"
    },
    {
      "affs": [
        3
      ],
      "name": "Randall T Peterson",
      "orcid": "0000-0003-0727-3469"
    }
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  "cited_by": 0,
  "doi": "10.1371/journal.pone.0355748",
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  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Biochemistry"
    ],
    [
      "heading",
      "Biology and Life Sciences"
    ],
    [
      "heading",
      "Biology and life sciences"
    ],
    [
      "heading",
      "Cell Biology"
    ],
    [
      "heading",
      "Cell Signaling"
    ],
    [
      "heading",
      "DNA construction"
    ],
    [
      "heading",
      "DNA-binding proteins"
    ],
    [
      "heading",
      "Electromagnetic Radiation"
    ],
    [
      "heading",
      "Enzymes"
    ],
    [
      "heading",
      "Enzymology"
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      "heading",
      "Genetic Loci"
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      "heading",
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      "heading",
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      "heading",
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    ],
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      "heading",
      "Guide RNA"
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      "heading",
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      "Luciferase"
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      "Luminescence"
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      "heading",
      "Molecular biology techniques"
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      "Research Article"
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    [
      "heading",
      "Research and analysis methods"
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      "heading",
      "Signal Transduction"
    ],
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      "plos",
      "/Biology and life sciences/Biochemistry/Enzymology/Enzymes/Oxidoreductases/Luciferase"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Nucleic acids/RNA/Guide RNA"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Proteins/DNA-binding proteins"
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    [
      "plos",
      "/Biology and life sciences/Biochemistry/Proteins/Enzymes/Oxidoreductases/Luciferase"
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    [
      "plos",
      "/Biology and life sciences/Cell biology/Signal transduction/Cell signaling/Genomic signal processing"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Genetic loci"
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    [
      "plos",
      "/Biology and life sciences/Genetics/Genomics/Human genomics"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/DNA construction/Plasmid construction"
    ],
    [
      "plos",
      "/Physical sciences/Physics/Electromagnetic radiation/Luminescence"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/DNA construction/Plasmid construction"
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  "pdf_url": "https://europepmc.org/articles/PMC13719016?pdf=render",
  "pmcid": "PMC13719016",
  "pmid": "42956069",
  "published": "2026-10-29",
  "publisher": "Public Library of Science",
  "ris": "https://research.draconitic.com/paper/1srmwhc0zmb0z.ris",
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  "subjects": [
    {
      "field": "biochemistry",
      "origin": "plos:enzymology",
      "subfield": "enzymology-metabolism"
    },
    {
      "field": "biochemistry",
      "origin": "heading:Molecular biology",
      "subfield": "molecular-biology"
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    {
      "field": "biology",
      "origin": "plos:cell biology",
      "subfield": "cell-biology"
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    {
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    {
      "field": "physics",
      "origin": "heading:Physics",
      "subfield": "general"
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  "title": "A dCas9 proximity reporter for investigating DNA linear and rotational dynamics",
  "title_text": "A dCas9 proximity reporter for investigating DNA linear and rotational dynamics",
  "type": "peer-reviewed",
  "updated_at": "2026-11-02T12:22:56Z",
  "url": "https://research.draconitic.com/paper/1srmwhc0zmb0z",
  "venue": "PLOS One",
  "versions": []
}