{
  "abstract": "Fluorescent proteins and small-molecule dyes offer complementary advantages for biological imaging: Proteins are amenable to genetic tagging, whereas dyes provide superior brightness and photostability. To combine these strengths, we used de novo protein design to generate small, nanomolar-affinity, high-selectivity binders (NovoTags) for three cell-permeable dyes spanning the visible spectrum. We show that the NovoTag fluorescent lifetimes can be tuned and demonstrate their application in lifetime- and wavelength-based multiplexed fluorescence imaging. We also designed a two-chain version (NovoSplit) that functions as a chemically induced dimerization system with fluorescent readout in living cells or as a minimally perturbing proximity probe in fixed cells. Our approach combines the advantages of fluorescent proteins and small-molecule dyes, thus expanding the toolkit for cellular imaging.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Chemical Engineering, University of Washington, Seattle, WA, USA",
    "Institute for Protein Design, University of Washington, Seattle, WA, USA",
    "Molecular Systems Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany",
    "Department of Biochemistry, University of Washington, Seattle, WA, USA",
    "Department of Chemistry, Stanford University, Stanford, CA, USA",
    "Graduate Program in Molecular Engineering, University of Washington, Seattle, WA, USA",
    "Department of Bioengineering, University of Washington, Seattle, WA, USA",
    "Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, South Korea",
    "Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA",
    "Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany",
    "Howard Hughes Medical Institute, University of Washington, Seattle, WA, USA"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 25,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Long Tran",
      "orcid": "0000-0001-9723-3529"
    },
    {
      "affs": [
        2
      ],
      "name": "Steffen Klein",
      "orcid": "0000-0002-8046-0474"
    },
    {
      "affs": [
        3,
        4,
        5,
        1
      ],
      "name": "David Juergens",
      "orcid": "0000-0001-6425-8391"
    },
    {
      "affs": [
        6,
        1
      ],
      "name": "Shajesh Sharma",
      "orcid": "0000-0001-9829-8631"
    },
    {
      "affs": [
        1
      ],
      "name": "Justin A. Decarreau",
      "orcid": "0000-0001-8996-7741"
    },
    {
      "affs": [
        3,
        7,
        1
      ],
      "name": "Gyu Rie Lee",
      "orcid": "0000-0002-9119-5303"
    },
    {
      "affs": [
        3,
        1
      ],
      "name": "Yujia Wang",
      "orcid": "0000-0002-7099-6283"
    },
    {
      "affs": [
        3,
        1
      ],
      "name": "Wei Chen",
      "orcid": "0000-0002-5255-4166"
    },
    {
      "affs": [
        1
      ],
      "name": "Asim K. Bera",
      "orcid": "0000-0001-9473-2912"
    },
    {
      "affs": [
        1
      ],
      "name": "Alex Kang",
      "orcid": "0000-0001-5487-0499"
    },
    {
      "affs": [
        1
      ],
      "name": "Jonathan Woods",
      "orcid": "0000-0002-5457-2464"
    },
    {
      "affs": [
        1
      ],
      "name": "Emily Joyce",
      "orcid": "0009-0007-3230-1341"
    },
    {
      "affs": [
        1
      ],
      "name": "Dionne K. Vafeados",
      "orcid": "0000-0002-2560-8907"
    },
    {
      "affs": [
        1
      ],
      "name": "Nicole Roullier",
      "orcid": "0009-0004-8430-9538"
    },
    {
      "affs": [
        1
      ],
      "name": "Xinting Li",
      "orcid": "0000-0001-5559-7578"
    },
    {
      "affs": [
        3,
        1
      ],
      "name": "Bingxu Liu"
    },
    {
      "affs": [
        3,
        1
      ],
      "name": "Bo Yang",
      "orcid": "0000-0001-5617-5748"
    },
    {
      "affs": [
        3,
        1
      ],
      "name": "Edin Muratspahić",
      "orcid": "0000-0002-6011-0805"
    },
    {
      "affs": [
        8
      ],
      "name": "Timothy A. Brown",
      "orcid": "0000-0001-8941-7678"
    },
    {
      "affs": [
        8
      ],
      "name": "Jonathan B. Grimm",
      "orcid": "0000-0003-0331-4200"
    },
    {
      "affs": [
        8
      ],
      "name": "Ronak H. Patel",
      "orcid": "0000-0002-6945-6466"
    },
    {
      "affs": [
        8
      ],
      "name": "Luke D. Lavis",
      "orcid": "0000-0002-0789-6343"
    },
    {
      "affs": [
        9,
        2
      ],
      "name": "Julia Mahamid",
      "orcid": "0000-0001-6968-041X"
    },
    {
      "affs": [
        3,
        1
      ],
      "name": "Linna An",
      "orcid": "0000-0001-9858-9105"
    },
    {
      "affs": [
        3,
        10,
        1
      ],
      "name": "David A. Baker",
      "orcid": "0000-0001-7896-6217"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/39jc5de3ad6jj.bib",
  "cited_by": 11,
  "doi": "10.1126/science.aeb0822",
  "fulltext_url": null,
  "id": "39jc5de3ad6jj",
  "issns": [
    "0036-8075",
    "0193-4511",
    "1095-9203"
  ],
  "landing_url": "https://doi.org/10.1126/science.aeb0822",
  "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": [
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      "openalex",
      "1304|Advanced Fluorescence Microscopy Techniques"
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  "openalex": "W7169058951",
  "pdf_url": null,
  "pmcid": "PMC12424662",
  "pmid": "40950233",
  "published": "2026-07-16",
  "publisher": "American Association for the Advancement of Science",
  "ris": "https://research.draconitic.com/paper/39jc5de3ad6jj.ris",
  "sources": [
    {
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      "url": "https://doi.org/10.1126/science.aeb0822"
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  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1304|Advanced Fluorescence Microscopy Techniques",
      "subfield": "biophysics"
    }
  ],
  "title": "De novo design of orthogonal far-red, orange, and green fluorophore-binding proteins for multiplexed imaging",
  "title_text": "De novo design of orthogonal far-red, orange, and green fluorophore-binding proteins for multiplexed imaging",
  "type": "peer-reviewed",
  "updated_at": "2026-11-08T13:20:43Z",
  "url": "https://research.draconitic.com/paper/39jc5de3ad6jj",
  "venue": "Science",
  "versions": []
}