{
  "abstract": "Insects and fungi are among the most destructive pests and pathogens affecting agriculture and public health. Although chemical pesticides have long been the primary means of control, their broad‑spectrum activity and widespread use have accelerated resistance in many systems. RNA interference (RNAi) has therefore emerged as a promising species‑specific alternative. RNAi is initiated by double‑stranded RNA (dsRNA), which triggers sequence‑directed degradation of target transcripts. However, RNAi efficiency varies markedly across species, influenced by nuclease activity, dsRNA length, and the cellular uptake route. We previously showed that long dsRNA uptake in Aedes aegypti depends on clathrin‑mediated endocytosis (CME), whereas paperclip RNA (pcRNA), a compact dsRNA structure developed in our laboratory, bypasses CME and enters cells through a distinct pathway. Here, we compared the uptake and RNAi performance of two dsRNA formats, long dsRNA (210–230 bp) and pcRNA (21–23 bp), in three representative species: Aedes aegypti (mosquito), Spodoptera frugiperda (moth), and Sclerotinia sclerotiorum (fungus). To assess cellular uptake of these dsRNAs, we used two CME inhibitors, chlorpromazine (CPZ) and bafilomycin A1 (BafA1), along with fluorescein‑labeled dsRNAs. CME inhibition prevented long dsRNA from entering the cytoplasm and abolished its ability to induce RNAi. In contrast, pcRNAs were readily internalized and produced significant gene knockdowns under both inhibitor treatments. Microscopy further revealed that all three cell types, pretreated with either CPZ or BafA1, prevented long dsRNA from entering the cytoplasm. pcRNA, consistently reached the cytoplasm and showed superior uptake across all species. Together, these findings demonstrate that pcRNA uses an uptake mechanism distinct from long dsRNA, enabling it to circumvent CME-dependent barriers. This unique internalization route likely reflects its compact size and structural configuration and highlights pcRNA as a promising platform for improving RNAi-based pest and pathogen control.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Biological Sciences, University of Manitoba, Winnipeg, Manitoba, Canada.",
    "Joint Genome Institute, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America."
  ],
  "article_type": "Research Article",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Nazanin Amanat",
      "orcid": "0000-0002-8797-446X"
    },
    {
      "affs": [
        1
      ],
      "name": "Bliss M Kernodle",
      "orcid": "0000-0002-2759-2566"
    },
    {
      "affs": [
        0
      ],
      "name": "Brigo Castillo",
      "orcid": "0000-0002-0939-9762"
    },
    {
      "affs": [
        0
      ],
      "name": "Alison Tayler"
    },
    {
      "affs": [
        0
      ],
      "name": "Mark Belmonte"
    },
    {
      "affs": [
        0
      ],
      "name": "Steve Whyard",
      "orcid": "0000-0002-9874-7780"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/58hpchxkwaetq.bib",
  "cited_by": 0,
  "doi": "10.1371/journal.pone.0359888",
  "fulltext_url": "https://europepmc.org/articles/PMC13716168",
  "id": "58hpchxkwaetq",
  "issns": [
    "1932-6203"
  ],
  "landing_url": "https://doi.org/10.1371/journal.pone.0359888",
  "language": "eng",
  "licence": "CC0-1.0",
  "licence_label": "CC0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/publicdomain/zero/1.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Aedes Aegypti"
    ],
    [
      "heading",
      "Animals"
    ],
    [
      "heading",
      "Arthropoda"
    ],
    [
      "heading",
      "Biochemistry"
    ],
    [
      "heading",
      "Biology and Life Sciences"
    ],
    [
      "heading",
      "Biology and life sciences"
    ],
    [
      "heading",
      "Cell Biology"
    ],
    [
      "heading",
      "Cellular Structures and Organelles"
    ],
    [
      "heading",
      "Cytoplasm"
    ],
    [
      "heading",
      "Disease Vectors"
    ],
    [
      "heading",
      "Entomology"
    ],
    [
      "heading",
      "Epigenetics"
    ],
    [
      "heading",
      "Eukaryota"
    ],
    [
      "heading",
      "Fluorescence Imaging"
    ],
    [
      "heading",
      "Fungal Pathogens"
    ],
    [
      "heading",
      "Fungal Structure"
    ],
    [
      "heading",
      "Gene expression"
    ],
    [
      "heading",
      "Gene regulation"
    ],
    [
      "heading",
      "Genetic interference"
    ],
    [
      "heading",
      "Genetics"
    ],
    [
      "heading",
      "Imaging Techniques"
    ],
    [
      "heading",
      "Infectious Diseases"
    ],
    [
      "heading",
      "Insect Vectors"
    ],
    [
      "heading",
      "Insects"
    ],
    [
      "heading",
      "Invertebrates"
    ],
    [
      "heading",
      "Medical Conditions"
    ],
    [
      "heading",
      "Medical Microbiology"
    ],
    [
      "heading",
      "Medicine and Health Sciences"
    ],
    [
      "heading",
      "Microbial Pathogens"
    ],
    [
      "heading",
      "Microbiology"
    ],
    [
      "heading",
      "Mosquitoes"
    ],
    [
      "heading",
      "Mycology"
    ],
    [
      "heading",
      "Non-coding RNA"
    ],
    [
      "heading",
      "Nucleic acids"
    ],
    [
      "heading",
      "Organisms"
    ],
    [
      "heading",
      "Pathogens"
    ],
    [
      "heading",
      "Pathology and Laboratory Medicine"
    ],
    [
      "heading",
      "RNA"
    ],
    [
      "heading",
      "RNA interference"
    ],
    [
      "heading",
      "Research Article"
    ],
    [
      "heading",
      "Research and Analysis Methods"
    ],
    [
      "heading",
      "Small interfering RNA"
    ],
    [
      "heading",
      "Species Interactions"
    ],
    [
      "heading",
      "Zoology"
    ],
    [
      "openalex",
      "1312|RNA Interference and Gene Delivery"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Nucleic acids/RNA/Non-coding RNA/Small interfering RNA"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Nucleic acids/RNA/RNA interference"
    ],
    [
      "plos",
      "/Biology and life sciences/Cell biology/Cellular structures and organelles/Cytoplasm"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Epigenetics/RNA interference"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Gene expression/Gene regulation/Small interfering RNA"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Gene expression/RNA interference"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Genetic interference/RNA interference"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Medical microbiology/Microbial pathogens/Fungal pathogens"
    ],
    [
      "plos",
      "/Biology and life sciences/Mycology/Fungal pathogens"
    ],
    [
      "plos",
      "/Biology and life sciences/Mycology/Fungal structure"
    ],
    [
      "plos",
      "/Biology and life sciences/Organisms/Eukaryota/Animals/Invertebrates/Arthropoda/Insects"
    ],
    [
      "plos",
      "/Biology and life sciences/Organisms/Eukaryota/Animals/Invertebrates/Arthropoda/Insects/Mosquitoes/Aedes aegypti"
    ],
    [
      "plos",
      "/Biology and life sciences/Species interactions/Disease vectors/Insect vectors/Mosquitoes/Aedes aegypti"
    ],
    [
      "plos",
      "/Biology and life sciences/Zoology/Animals/Invertebrates/Arthropoda/Insects"
    ],
    [
      "plos",
      "/Biology and life sciences/Zoology/Animals/Invertebrates/Arthropoda/Insects/Mosquitoes/Aedes aegypti"
    ],
    [
      "plos",
      "/Biology and life sciences/Zoology/Entomology/Insects"
    ],
    [
      "plos",
      "/Biology and life sciences/Zoology/Entomology/Insects/Mosquitoes/Aedes aegypti"
    ],
    [
      "plos",
      "/Medicine and health sciences/Medical conditions/Infectious diseases/Disease vectors/Insect vectors/Mosquitoes/Aedes aegypti"
    ],
    [
      "plos",
      "/Medicine and health sciences/Pathology and laboratory medicine/Pathogens/Microbial pathogens/Fungal pathogens"
    ],
    [
      "plos",
      "/Research and analysis methods/Imaging techniques/Fluorescence imaging"
    ]
  ],
  "openalex": "W7224079689",
  "pdf_url": "https://europepmc.org/articles/PMC13716168?pdf=render",
  "pmcid": "PMC13716168",
  "pmid": "42949895",
  "published": "2026-10-28",
  "publisher": "Public Library of Science",
  "ris": "https://research.draconitic.com/paper/58hpchxkwaetq.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:17Z",
      "native_id": "10.1371/journal.pone.0359888",
      "source": "plos",
      "url": "https://doi.org/10.1371/journal.pone.0359888"
    },
    {
      "first_seen": "2026-10-31T00:20:33Z",
      "native_id": "MED:42949895",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42949895"
    },
    {
      "first_seen": "2026-11-01T12:23:13Z",
      "native_id": "PMC13716168",
      "source": "pmc",
      "url": "https://doi.org/10.1371/journal.pone.0359888"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "heading:Gene expression",
      "subfield": "molecular-biology"
    },
    {
      "field": "biology",
      "origin": "heading:Cell Biology",
      "subfield": "cell-biology"
    },
    {
      "field": "biology",
      "origin": "plos:genetics",
      "subfield": "genetics-genomics"
    },
    {
      "field": "biology",
      "origin": "plos:microbiology",
      "subfield": "microbiology"
    },
    {
      "field": "biology",
      "origin": "plos:zoology",
      "subfield": "zoology-behaviour"
    }
  ],
  "title": "Paperclip RNAs bypass clathrin mediated endocytosis in insect and fungal cells",
  "title_text": "Paperclip RNAs bypass clathrin mediated endocytosis in insect and fungal cells",
  "type": "peer-reviewed",
  "updated_at": "2026-11-01T12:23:13Z",
  "url": "https://research.draconitic.com/paper/58hpchxkwaetq",
  "venue": "PLOS One",
  "versions": []
}