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PreprintCited by 0Advanced Fluorescence Microscopy TechniquesGenetics, Aging, and Longevity in Model OrganismsDevelopmental Biology and Gene Regulation

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A pilot study for whole proteome tagging in C. elegans

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Eroglu, Matthew, Hobert, Oliver·2026·eLife
DOI 10.64898/2026.02.09.704846PMC PMC12919041

Preprint — published as 10.7554/elife.110717.4 · Paper page

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Abstract

Tagging all proteins encoded by an animal genome with a fluorescent tag would open many windows to the discovery of unexpected patterns of protein expression and localization. To scale such an approach, it would be beneficial to introduce multiple, spectrally distinct fluorophore tags in parallel. As proof of concept for scalable pooled tagging, we undertook a pilot study in the nematode C. elegans, in which we set out to tag 30 different genetic loci with three different fluorophores, with 3 tags being introduced at a time. By choosing essential genes, predicted based on transcriptomics to cover a range of expression levels, we explore issues relating to disrupting gene function and visibility of tagged proteins. We demonstrate that such a tagging approach is highly efficient and indeed reveals unanticipated patterns of cellular sites of expression, as well as subcellular protein localization. We hope that this pilot study will motivate attempts to scale this tagging approach to more loci and, ultimately, the whole genome.

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Metadata

Authors
Eroglu, M., & Hobert, O.
Affiliations
Howard Hughes Medical Institute (US)
Corresponding author
Hobert, Oliver
ORCID
Venue
eLife
Publisher
openRxiv
Published
2026-02-10
Volume
15
ISSN
2050-084X (ISSN-L)
DOI
10.64898/2026.02.09.704846
PMC
PMC12919041
OpenAlex
W7128481866
Cited by
0
References
65
Type
posted content
Language
en
Open access
Licence
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Howard Hughes Medical Institute
  • Leon Levy Foundation
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
Keywords
fluorescent protein tags, Caenorhabditis elegans, essential genes, protein expression patterns, subcellular protein localization

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