{
  "abstract": "The design of RNA-guided nucleases with properties not limited by evolution can expand programmable genome-editing capabilities. However, generating diverse multidomain proteins with robust enzymatic properties remains challenging. Here, we use a protein design strategy that couples a structure-guided inverse-folding model with evolution-informed residue constraints to generate active, divergent variants of TnpB, a minimal CRISPR-Cas12-like nuclease, termed SynTnpBs. High-throughput screening of artificial intelligence-generated variants yielded editors that retained or exceeded wild-type activity in bacterial, plant, and human cells. Cryo-electron microscopy-based structure determination of the most divergent variant revealed stabilizing contacts in the RNA-DNA interfaces across conformations, demonstrating the design potential of this approach. Together, these results establish a strategy for creating non-natural RNA-guided nucleases and conformationally active nucleic acid binders, enlarging the designable protein space.",
  "abstract_shown": true,
  "affiliations": [
    "California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, CA, USA",
    "Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA",
    "Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA",
    "Department of Molecular, Cell and Developmental Biology, University of California at Los Angeles, Los Angeles, CA, USA",
    "Biophysics Graduate Group, University of California, Berkeley, Berkeley, CA, CA, USA",
    "Center for Computational Biology, University of California Berkeley, Berkeley, CA, USA",
    "Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA",
    "Department of Earth and Planetary Science, University of California, Berkeley, CA, USA",
    "Department of Microbiology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia",
    "Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA",
    "Howard Hughes Medical Institute (HHMI), University of California at Los Angeles, Los Angeles, CA, USA",
    "Gladstone Institute of Data Science and Biotechnology, San Francisco, CA, USA",
    "Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA",
    "Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA, USA",
    "Li Ka Shing Center for Genomic Engineering, University of California, Berkeley, Berkeley, CA, USA"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 20,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Petr Skopintsev",
      "orcid": "0000-0002-6043-157X"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Isabel Esain-Garcia",
      "orcid": "0000-0003-1931-4563"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Evan C. DeTurk",
      "orcid": "0009-0004-9613-8625"
    },
    {
      "affs": [
        2,
        1
      ],
      "name": "Peter H. Yoon",
      "orcid": "0000-0002-9156-1393"
    },
    {
      "affs": [
        2,
        1
      ],
      "name": "Zehan Zhou"
    },
    {
      "affs": [
        3
      ],
      "name": "Trevor J. Weiss",
      "orcid": "0000-0002-2373-0802"
    },
    {
      "affs": [
        3
      ],
      "name": "Maris Kamalu",
      "orcid": "0009-0009-8008-7196"
    },
    {
      "affs": [
        2,
        1
      ],
      "name": "Ajit Chamraj",
      "orcid": "0009-0005-7114-1753"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Kenneth J. Loi",
      "orcid": "0009-0009-5508-6511"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Conner J. Langeberg",
      "orcid": "0000-0002-5609-3758"
    },
    {
      "affs": [
        4,
        0,
        1
      ],
      "name": "Ron S. Boger",
      "orcid": "0000-0002-4467-271X"
    },
    {
      "affs": [
        5
      ],
      "name": "Hunter M. Nisonoff",
      "orcid": "0000-0003-1357-8111"
    },
    {
      "affs": [
        6,
        1
      ],
      "name": "Hannah Karp",
      "orcid": "0000-0002-1619-5901"
    },
    {
      "affs": [
        7
      ],
      "name": "Lin-Xing Chen",
      "orcid": "0000-0003-2774-1952"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Honglue Shi",
      "orcid": "0000-0003-3847-1652"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Kamakshi Vohra",
      "orcid": "0009-0004-4368-2694"
    },
    {
      "affs": [
        7,
        8,
        1
      ],
      "name": "Jillian F. Banfield",
      "orcid": "0000-0001-8203-8771"
    },
    {
      "affs": [
        4,
        0,
        2,
        1,
        9
      ],
      "name": "Jamie H. D. Cate",
      "orcid": "0000-0001-5965-7902"
    },
    {
      "affs": [
        3,
        10
      ],
      "name": "Steven E. Jacobsen",
      "orcid": "0000-0001-9483-138X"
    },
    {
      "affs": [
        4,
        0,
        2,
        11,
        12,
        13,
        1,
        14,
        9
      ],
      "name": "Jennifer A. Doudna",
      "orcid": "0000-0001-9161-999X"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/fsj8p9hbxk5ta.bib",
  "cited_by": 7,
  "doi": "10.1126/science.aed6123",
  "fulltext_url": null,
  "id": "fsj8p9hbxk5ta",
  "issns": [
    "0036-8075",
    "0193-4511",
    "1095-9203"
  ],
  "landing_url": "https://doi.org/10.1126/science.aed6123",
  "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"
    ]
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  "openalex": "W4417145943",
  "pdf_url": null,
  "pmcid": "PMC12822741",
  "pmid": "41573840",
  "published": "2026-07-16",
  "publisher": "American Association for the Advancement of Science",
  "ris": "https://research.draconitic.com/paper/fsj8p9hbxk5ta.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T13:26:48Z",
      "native_id": "W4417145943",
      "source": "openalex",
      "url": "https://doi.org/10.1126/science.aed6123"
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  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|CRISPR and Genetic Engineering",
      "subfield": "molecular-biology"
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  ],
  "title": "Structure and evolution-guided design of minimal RNA-guided nucleases",
  "title_text": "Structure and evolution-guided design of minimal RNA-guided nucleases",
  "type": "peer-reviewed",
  "updated_at": "2026-11-03T13:20:30Z",
  "url": "https://research.draconitic.com/paper/fsj8p9hbxk5ta",
  "venue": "Science",
  "versions": []
}