{
  "abstract": "LINE-1 (L1) is transcribed by RNA polymerase (Pol) II, whereas the non-coding Short INterspersed Element (SINE) Alu is transcribed by Pol III and exploits L1 ORF2p for mobilization. Conventional L1 retrotransposition reporters rely on spliceosome-dependent introns and cannot directly monitor Alu retrotransposition. Here, we developed EGFPTet, a fluorescent SINE reporter in which EGFP is interrupted by a Tetrahymena thermophila group I self-splicing intron. EGFPTet enabled flow cytometry, fluorescence microscopy, cell sorting, and time-course analysis of Alu mobilization without colony formation. Reporter activation required ORF2p reverse transcriptase activity, and recovered Alu-EGFPTet insertions displayed retrotransposition hallmarks. EGFPTet quantified Alu retrotransposition across cell lines and host-factor perturbations and was repurposed to detect mobilization of the phylogenetically distinct mouse B2 SINE. Defined amino acid substitutions generated an mTurquoise2 derivative without redesigning the self-splicing junctions, allowing dual-color analysis of Alu trans-mobilization and L1 cis retrotransposition in individual cells. In this assay, wild-type L1 supported a higher Alu retrotransposition frequency than the ORF1p-mutant L1. Despite their shared dependence on ORF2p, Alu co-expression did not significantly reduce L1 retrotransposition, and Alu/L1 dual-positive counts exceeded independence-based expectations, indicating preferential co-occurrence of the two reporter signals. EGFPTet provides a fluorescent platform for investigating the shared and distinct cellular conditions that support SINE and L1 mobilization.",
  "abstract_shown": true,
  "affiliations": [
    "RIKEN"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 4,
  "authors": [
    {
      "name": "S. Amano"
    },
    {
      "name": "K. Ohnishi"
    },
    {
      "name": "K. Nishimori"
    },
    {
      "affs": [
        0
      ],
      "name": "T. Miyoshi"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/88z7rm01mg6cf.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.09.26.754642",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.09.26.754642v1.full",
  "id": "88z7rm01mg6cf",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.09.26.754642v1",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "molecular biology"
    ],
    [
      "openalex",
      "1110|Chromosomal and Genetic Variations"
    ]
  ],
  "openalex": "W7214681622",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.09.26.754642v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-26",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/88z7rm01mg6cf.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:24:51Z",
      "native_id": "10.64898/2026.09.26.754642",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.09.26.754642v1"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "biorxiv:molecular biology",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Dual-color fluorescent reporters resolve Alu and LINE-1 retrotransposition in single cells",
  "title_text": "Dual-color fluorescent reporters resolve Alu and LINE-1 retrotransposition in single cells",
  "type": "preprint",
  "updated_at": "2026-10-30T12:24:51Z",
  "url": "https://research.draconitic.com/paper/88z7rm01mg6cf",
  "venue": "bioRxiv",
  "versions": []
}