{
  "abstract": "Residual gene editors are a concern associated with the widespread application of CRISPR/Cas9 technology. Here, we developed a CRISPR/Cas12a-based fluorescent aptasensor for the qualitative and quantitative detection of Cas9 protein. Secondary structure prediction and molecular docking were combined to identify potential binding sites of the Cas9-specific aptamer. A fluorescence aptasensor was constructed based on competitive binding: in the presence of Cas9, the aptamer binds to the target and releases complementary DNA (cDNA), which activates the trans-cleavage activity of Cas12a. Cas12a subsequently cleaves a single-stranded DNA (ssDNA) reporter labeled with a fluorophore and a quencher, generating a fluorescence signal, expressed in relative fluorescence units (RFU), that increases with Cas9 concentration. The optimized conditions comprised a trans-cleavage reaction time of 20 min, a Cas12a:CRISPR RNA (crRNA) molar ratio of 1:1, a final aptamer concentration of 180 nM, and a combination of low- and high-salt buffers to balance molecular recognition and protein stability. The proposed assay enabled Cas9 protein detection in buffer and Cas9-spiked soybean milk and cereal matrices. These results represent a proof-of-concept evaluation in complex food matrices rather than validation of authentic residual Cas9 detection. Further studies using unspiked gene-edited samples with independently verified Cas9 residues and an orthogonal reference method are required.",
  "abstract_shown": true,
  "affiliations": [
    "College of Life Science and Technology, Beijing University of Chemical Technology, Chaoyang District, Beijing 100029, China",
    "Quality and Safety Institute of Agricultural Products, Heilongjiang Academy of Agricultural Sciences/Key Laboratory of Quality and Safety of Cereals and Their Products, Harbin 150086, China",
    "Hangzhou Institute of Medicine (HIM), The Chinese Academy of Sciences, Hangzhou 310022, China"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 7,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Zizhao Wang"
    },
    {
      "affs": [
        1
      ],
      "name": "Jing Wang",
      "orcid": "0000-0003-0444-4744"
    },
    {
      "affs": [
        2
      ],
      "name": "Ling Li"
    },
    {
      "affs": [
        1
      ],
      "name": "Guofeng Chen"
    },
    {
      "affs": [
        1
      ],
      "name": "Hongtao Wen",
      "orcid": "0000-0001-7374-3073"
    },
    {
      "affs": [
        1
      ],
      "name": "Baohai Liu",
      "orcid": "0009-0008-9347-6628"
    },
    {
      "affs": [
        0
      ],
      "name": "Yiyang Dong",
      "orcid": "0000-0003-1752-3935"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/7xtjjd7w85b6t.bib",
  "cited_by": 0,
  "doi": "10.3390/molecules31203607",
  "fulltext_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13707788/?tool=EBI",
  "id": "7xtjjd7w85b6t",
  "issns": [
    "1420-3049"
  ],
  "landing_url": "https://www.mdpi.com/1420-3049/31/20/3607",
  "language": "eng",
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Advanced biosensing and bioanalysis techniques"
    ]
  ],
  "openalex": "W7221330495",
  "pdf_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13707788/pdf/?tool=EBI",
  "pmcid": "PMC13707788",
  "pmid": null,
  "published": "2026-10-01",
  "publisher": "MDPI AG",
  "ris": "https://research.draconitic.com/paper/7xtjjd7w85b6t.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:18:42Z",
      "native_id": "10.3390/molecules31203607",
      "source": "crossref",
      "url": "https://www.mdpi.com/1420-3049/31/20/3607"
    },
    {
      "first_seen": "2026-10-30T12:21:34Z",
      "native_id": "PMC:PMC13707788",
      "source": "europepmc",
      "url": "https://europepmc.org/article/PMC/PMC13707788"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Advanced biosensing and bioanalysis techniques",
      "subfield": "molecular-biology"
    }
  ],
  "title": "CRISPR/Cas12a-Based Aptasensor for Cas9 Protein Detection in Spiked Food Matrices",
  "title_text": "CRISPR/Cas12a-Based Aptasensor for Cas9 Protein Detection in Spiked Food Matrices",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:21:34Z",
  "url": "https://research.draconitic.com/paper/7xtjjd7w85b6t",
  "venue": "Molecules",
  "versions": []
}