{
  "abstract": "Precise, site-specific integration of large DNA sequences into plant genomes is a long-standing goal of crop biotechnology and synthetic biology, yet remains constrained by random insertion, inefficient homologous recombination, and gene targeting. Here, we present PrimeStack, a DSB-independent platform that integrates PE with the unidirectional large serine integrase Bxb1, leveraging evolved variants for enhanced activity, to achieve the programmable insertion of multigene, multikilobase cargos at predefined genomic safe harbors (GSH) in rice (Oryza sativa). Optimized prime editors first install attP landing sites with high fidelity and heritability followed by Bxb1-mediated recombination that generates irreversible integration of genetic information. PrimeStack enabled targeted large-cargo integration in regenerated rice plants with efficiencies of up to 45%, as determined by junction-specific PCR analyses. No obvious morphological differences from WT plants were observed at 5 weeks after transfer to soil under the evaluated conditions. We validate its utility by achieving targeted insertion of a carotenoid cassette at an experimentally confirmed genomic safe harbor. PrimeStack delivers a modular, site-specific gene stacking platform that enables predictable, multigene trait pyramiding and pathway construction at GSH, thereby accelerating the development of improved and resilient crop varieties, as well as scalable plant-based biomanufacturing and a powerful chassis for synthetic biology.\n\nPrecise integration of large DNA sequences into plant genomes remains a major challenge. This study presents PrimeStack, a platform that combines prime editing with the Bxb1 integrase to achieve the programmable insertion of multigene cargos at plant genomic safe harbors with high efficiency.",
  "abstract_shown": true,
  "affiliations": [
    "Laboratory for Genome Engineering and Synthetic Biology, Division of Biomedical Sciences, King Abdullah University of Science and Technology (KAUST), Thuwal, Saudi Arabia."
  ],
  "article_type": "Methods and Resources",
  "arxiv": null,
  "author_total": 4,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Edith Sánchez"
    },
    {
      "affs": [
        0
      ],
      "name": "Khalid Sedeek",
      "orcid": "0000-0002-2913-6628"
    },
    {
      "affs": [
        0
      ],
      "name": "Haroon Butt",
      "orcid": "0000-0001-6935-8025"
    },
    {
      "affs": [
        0
      ],
      "name": "Magdy Mahfouz",
      "orcid": "0000-0002-0616-6365"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/a4jynydeyztmt.bib",
  "cited_by": 0,
  "doi": "10.1371/journal.pbio.3003778",
  "fulltext_url": "https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3003778",
  "id": "a4jynydeyztmt",
  "issns": [
    "1545-7885",
    "1544-9173"
  ],
  "landing_url": "https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3003778",
  "language": "eng",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|CRISPR and Genetic Engineering"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Enzymology/Enzymes/Recombinases"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Proteins/Enzymes/Recombinases"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Genetic loci"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Genomics"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Heredity/Homozygosity"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Artificial gene amplification and extension/Polymerase chain reaction"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Sequencing techniques/DNA sequencing/Dideoxy DNA sequencing"
    ],
    [
      "plos",
      "/Biology and life sciences/Organisms/Eukaryota/Plants/Grasses/Rice"
    ],
    [
      "plos",
      "/Physical sciences/Materials science/Materials/Pigments/Organic pigments/Carotenoids"
    ],
    [
      "plos",
      "/Research and analysis methods/Animal studies/Experimental organism systems/Plant and algal models/Rice"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Artificial gene amplification and extension/Polymerase chain reaction"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Sequencing techniques/DNA sequencing/Dideoxy DNA sequencing"
    ]
  ],
  "openalex": "W7221584110",
  "pdf_url": "https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.3003778&type=printable",
  "pmcid": "PMC13693989",
  "pmid": "42868167",
  "published": "2026-10-13",
  "publisher": "Public Library of Science",
  "ris": "https://research.draconitic.com/paper/a4jynydeyztmt.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:30Z",
      "native_id": "10.1371/journal.pbio.3003778",
      "source": "plos",
      "url": "https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3003778"
    },
    {
      "first_seen": "2026-10-30T12:19:00Z",
      "native_id": "MED:42868167",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42868167"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "plos:molecular biology",
      "subfield": "molecular-biology"
    },
    {
      "field": "biology",
      "origin": "plos:genetics",
      "subfield": "genetics-genomics"
    }
  ],
  "title": "PrimeStack editors enable efficient and site-specific integration of large multigene cargos in plants",
  "title_text": "PrimeStack editors enable efficient and site-specific integration of large multigene cargos in plants",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:19:00Z",
  "url": "https://research.draconitic.com/paper/a4jynydeyztmt",
  "venue": "PLOS Biology",
  "versions": []
}