{
  "abstract": "CRISPR-Cas systems have emerged as powerful biosensing platforms capable of highly sensitive and specific nucleic acid detection. The performance of these assays critically depends on the design of the guide RNA (gRNA), which governs target recognition and activates cis - and trans -cleavage. However, native gRNAs often exhibit limited stability, weak mismatch discrimination, reduced activity in clinical matrices, and inconsistent collateral cleavage, thereby limiting the diagnostic potential of CRISPR sensors. To address these constraints, a wide spectrum of gRNA engineering strategies has been developed, including length change, structural reconfiguration, DNA modified strategies, chemical modifications, mismatch as well as multiple modifications. These modifications substantially enhance sensitivity, specificity, stability, and programmability across Cas12a, Cas13a, and emerging CRISPR effectors. This review systematically summarizes recent advances, compares mechanistic principles and functional outcomes, and highlights future opportunities for rational gRNA engineering to enable amplification-free detection, improve clinical robustness, and drive the development of next-generation CRISPR diagnostics.",
  "abstract_shown": true,
  "affiliations": [
    "School of Biomedical Engineering, UNSW Sydney, Sydney 2052, Australia",
    "Institute for Biomedical Materials and Devices (IBMD), School of Mathematical and Physical Sciences, Faculty of Science, University of Technology Sydney, Sydney, New South Wales, Australia",
    "State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China",
    "Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117599",
    "Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, 117558",
    "NUS Synthetic Biology for Clinical and Technological Innovation (SynCTI), National University of Singapore, 117456, Singapore",
    "National Centre for Engineering Biology (NCEB), Singapore, 117575",
    "Synthetic Biology Translational Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117456"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 7,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Wenyu Fu"
    },
    {
      "affs": [
        0
      ],
      "name": "Rui Sang",
      "orcid": "0000-0001-8557-2072"
    },
    {
      "affs": [
        0
      ],
      "name": "Ruier Xue",
      "orcid": "0009-0007-9956-1981"
    },
    {
      "affs": [
        1
      ],
      "name": "Guochen Bao",
      "orcid": "0000-0001-5103-5009"
    },
    {
      "affs": [
        2
      ],
      "name": "Shilun Feng",
      "orcid": "0000-0002-2560-2417"
    },
    {
      "affs": [
        3,
        4,
        5,
        6,
        7
      ],
      "name": "Chunlei Jiao"
    },
    {
      "affs": [
        0
      ],
      "name": "Fei Deng",
      "orcid": "0000-0002-5606-0278"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/az6n67q7v14a5.bib",
  "cited_by": 6,
  "doi": "10.1016/j.trac.2026.118960",
  "fulltext_url": null,
  "id": "az6n67q7v14a5",
  "issns": [
    "0165-9936",
    "0167-2940",
    "1879-3142"
  ],
  "landing_url": "https://doi.org/10.1016/j.trac.2026.118960",
  "language": "en",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|CRISPR and Genetic Engineering"
    ]
  ],
  "openalex": "W7163712950",
  "pdf_url": null,
  "pmcid": null,
  "pmid": null,
  "published": "2026-06-05",
  "publisher": "Elsevier BV",
  "ris": "https://research.draconitic.com/paper/az6n67q7v14a5.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T13:26:48Z",
      "native_id": "W7163712950",
      "source": "openalex",
      "url": "https://doi.org/10.1016/j.trac.2026.118960"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|CRISPR and Genetic Engineering",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Engineering gRNA for CRISPR biosensing: chemical, structural, and programmable designs for ultrasensitive diagnostics",
  "title_text": "Engineering gRNA for CRISPR biosensing: chemical, structural, and programmable designs for ultrasensitive diagnostics",
  "type": "peer-reviewed",
  "updated_at": "2026-11-01T13:20:13Z",
  "url": "https://research.draconitic.com/paper/az6n67q7v14a5",
  "venue": "TrAC Trends in Analytical Chemistry",
  "versions": []
}