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

The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.

A pilot study for whole proteome tagging in Caenorhabditis elegans

Open access

Published version of a preprint

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 Caenorhabditis elegans, in which we set out to tag 30 different genetic loci with three different fluorophores, with three 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 and subcellular sites of protein expression and localization. We hope that this pilot study will motivate attempts to scale this tagging approach to more loci and, ultimately, the whole genome.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.7554/elife.110717.4

Cites

3 of 3 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 58

  1. Adler, 2014, High fitness costs and instability of gene duplications reduce rates of evolution of new genes by duplication-divergence mechanisms, Molecular Biology and Evolution, 31, 1526, 10.1093/molbev/msu111 · DOI
  2. Altun, 2002, WormAtlas
  3. Ando, 2024, StayGold variants for molecular fusion and membrane-targeting applications, Nature Methods, 21, 648, 10.1038/s41592-023-02085-6 · DOI
  4. Arribere, 2014, Efficient marker-free recovery of custom genetic modifications with CRISPR/Cas9 in Caenorhabditis elegans, Genetics, 198, 837, 10.1534/genetics.114.169730 · DOI
  5. Bayer, 2025, The DEG/ENaC channel DEGT-1 is a proprioceptor of C. elegans foregut movement, Current Biology, 35, 5332, 10.1016/j.cub.2025.09.065 · DOI
  6. Brenner, 1974, The genetics of Caenorhabditis elegans, Genetics, 77, 71, 10.1093/genetics/77.1.71 · DOI
  7. Casper, 2026, The UCSC genome browser database: 2026 update, Nucleic Acids Research, 54, D1331, 10.1093/nar/gkaf1250 · DOI
  8. Dickinson, 2013, Engineering the Caenorhabditis elegans genome using Cas9-triggered homologous recombination, Nature Methods, 10, 1028, 10.1038/nmeth.2641 · DOI
  9. Eroglu, 2023, Efficient CRISPR/Cas9 mediated large insertions using long single-stranded oligonucleotide donors in C. elegans, The FEBS Journal, 290, 4429, 10.1111/febs.16876 · DOI
  10. Eroglu, 2024, Noncanonical inheritance of phenotypic information by protein amyloids, Nature Cell Biology, 26, 1712, 10.1038/s41556-024-01494-9 · DOI
  11. Gadella, 2023, mScarlet3: a brilliant and fast-maturing red fluorescent protein, Nature Methods, 20, 541, 10.1038/s41592-023-01809-y · DOI
  12. Gao, 2024, Aging atlas reveals cell-type-specific effects of pro-longevity strategies, Nature Aging, 4, 998, 10.1038/s43587-024-00631-1 · DOI
  13. Gerstein, 2010, Integrative analysis of the Caenorhabditis elegans genome by the modENCODE project, Science, 330, 1775, 10.1126/science.1196914 · DOI
  14. Ghaddar, 2023, Whole-body gene expression atlas of an adult metazoan, Science Advances, 9, 10.1126/sciadv.adg0506 · DOI
  15. Ghanta, 2020, Melting dsDNA donor molecules greatly improves precision genome editing in Caenorhabditis elegans, Genetics, 216, 643, 10.1534/genetics.120.303564 · DOI
  16. Götzke, 2019, The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications, Nature Communications, 10, 10.1038/s41467-019-12301-7 · DOI
  17. Grün, 2014, Conservation of mRNA and protein expression during development of C. elegans, Cell Reports, 6, 565, 10.1016/j.celrep.2014.01.001 · DOI
  18. Guo, 2025, Mitochondrial proteome reveals metabolic tuning by restricted insulin signaling to promote longevity in Caenorhabditis elegans, Biology, 14, 10.3390/biology14030279 · DOI
  19. Harris, 2020, WormBase: a modern model organism information resource, Nucleic Acids Research, 48, D762, 10.1093/nar/gkz920 · DOI
  20. Hu, 2011, A neuropeptide-mediated stretch response links muscle contraction to changes in neurotransmitter release, Neuron, 71, 92, 10.1016/j.neuron.2011.04.021 · DOI
  21. Huff, 2015, The Airyscan detector from ZEISS: confocal imaging with improved signal-to-noise ratio and super-resolution, Nature Methods, 12, i, 10.1038/nmeth.f.388 · DOI
  22. Huh, 2003, Global analysis of protein localization in budding yeast, Nature, 425, 686, 10.1038/nature02026 · DOI · Paper page
  23. Keeley, 2020, Comprehensive endogenous tagging of basement membrane components reveals dynamic movement within the matrix scaffolding, Developmental Cell, 54, 60, 10.1016/j.devcel.2020.05.022 · DOI
  24. Kumar, 2014, Dual-view plane illumination microscopy for rapid and spatially isotropic imaging, Nature Protocols, 9, 2555, 10.1038/nprot.2014.172 · DOI
  25. Lambert, 2019, FPbase: a community-editable fluorescent protein database, Nature Methods, 16, 277, 10.1038/s41592-019-0352-8 · DOI
  26. Leyhr, 2025, WormTagDB: a systematic survey of endogenously tagged proteins in C. elegans and roadmap towards the tagged proteome, 10.64898/2025.12.02.691955 · DOI
  27. Leyva-Díaz, 2025, Alternative splicing controls pan-neuronal homeobox gene expression, Genes & Development, 39, 209, 10.1101/gad.352184.124 · DOI
  28. Li, 2009, Proteomic analysis of mitochondria from Caenorhabditis elegans, PROTEOMICS, 9, 4539, 10.1002/pmic.200900101 · DOI
  29. Liu, 2016, On the dependency of cellular protein levels on mRNA abundance, Cell, 165, 535, 10.1016/j.cell.2016.03.014 · DOI · Paper page
  30. Lynch, 2000, The evolutionary fate and consequences of duplicate genes, Science, 290, 1151, 10.1126/science.290.5494.1151 · DOI · Paper page
  31. Mawuenyega, 2003, Large-scale identification of Caenorhabditis elegans proteins by multidimensional liquid chromatography-tandem mass spectrometry, Journal of Proteome Research, 2, 23, 10.1021/pr025551y · DOI
  32. Paix, 2015, High efficiency, homology-directed genome editing in Caenorhabditis elegans using CRISPR-Cas9 ribonucleoprotein complexes, Genetics, 201, 47, 10.1534/genetics.115.179382 · DOI
  33. Palmer, 2004, Investigation into the use of C- and N-terminal GFP fusion proteins for subcellular localization studies using reverse transfection microarrays, Comparative and Functional Genomics, 5, 342, 10.1002/cfg.405 · DOI
  34. Pan, 2024, Robotic microinjection enables large-scale transgenic studies of Caenorhabditis elegans, Nature Communications, 15, 10.1038/s41467-024-53108-5 · DOI
  35. Preibisch, 2009, Globally optimal stitching of tiled 3D microscopic image acquisitions, Bioinformatics, 25, 1463, 10.1093/bioinformatics/btp184 · DOI · Paper page
  36. Ragle, 2025, Multiscale patterning of a model apical extracellular matrix revealed by systematic endogenous protein tagging, 10.1101/2025.05.14.653803 · DOI
  37. Rankin, 2011, Nanoscopy in a living multicellular organism expressing GFP, Biophysical Journal, 100, L63, 10.1016/j.bpj.2011.05.020 · DOI
  38. Reilly, 2020, Unique homeobox codes delineate all the neuron classes of C. elegans, Nature, 584, 595, 10.1038/s41586-020-2618-9 · DOI
  39. Reilly, 2022, Widespread employment of conserved C. elegans homeobox genes in neuronal identity specification, PLOS Genetics, 18, 10.1371/journal.pgen.1010372 · DOI
  40. Ryu, 2024, Cellular ATP demand creates metabolically distinct subpopulations of mitochondria, Nature, 635, 746, 10.1038/s41586-024-08146-w · DOI · Paper page
  41. Santella, 2022, Cross-modality synthesis of EM time series and live fluorescence imaging, eLife, 11, 10.7554/eLife.77918 · DOI
  42. Schindelin, 2012, Fiji: an open-source platform for biological-image analysis, Nature Methods, 9, 676, 10.1038/nmeth.2019 · DOI · Paper page
  43. Simmer, 2003, Genome-wide RNAi of C. elegans using the hypersensitive rrf-3 strain reveals novel gene functions, PLOS Biology, 1, 10.1371/journal.pbio.0000012 · DOI
  44. Spieth, 2014, Overview of gene structure in C. elegans, WormBook, 1, 1, 10.1895/wormbook.1.65.2 · DOI
  45. Subach, 2011, An enhanced monomeric blue fluorescent protein with the high chemical stability of the chromophore, PLOS ONE, 6, 10.1371/journal.pone.0028674 · DOI
  46. Suh, 2012, A survey of putative secreted and transmembrane proteins encoded in the C. elegans genome, BMC Genomics, 13, 10.1186/1471-2164-13-333 · DOI
  47. Sural, 2025, Gut epithelium modifies enteric behaviors during nutritional adversity via distinct peptidergic signaling axes, Science Advances, 11, 10.1126/sciadv.adw1270 · DOI
  48. Taylor, 2021, Molecular topography of an entire nervous system, Cell, 184, 4329, 10.1016/j.cell.2021.06.023 · DOI
  49. Toker, 2025, Divergence in neuronal signaling pathways despite conserved neuronal identity among Caenorhabditis species, Current Biology, 35, 2927, 10.1016/j.cub.2025.05.036 · DOI
  50. Tomioka, 2006, The insulin/PI 3-kinase pathway regulates salt chemotaxis learning in Caenorhabditis elegans, Neuron, 51, 613, 10.1016/j.neuron.2006.07.024 · DOI
  51. Vergara, 2021, Whole-body integration of gene expression and single-cell morphology, Cell, 184, 4819, 10.1016/j.cell.2021.07.017 · DOI
  52. Vogel, 2012, Insights into the regulation of protein abundance from proteomic and transcriptomic analyses, Nature Reviews. Genetics, 13, 227, 10.1038/nrg3185 · DOI · Paper page
  53. Weill, 2019, Assessment of GFP tag position on protein localization and growth fitness in yeast, Journal of Molecular Biology, 431, 636, 10.1016/j.jmb.2018.12.004 · DOI
  54. Witvliet, 2021, Connectomes across development reveal principles of brain maturation, Nature, 596, 257, 10.1038/s41586-021-03778-8 · DOI · Paper page
  55. Wu, 2017, Reflective imaging improves spatiotemporal resolution and collection efficiency in light sheet microscopy, Nature Communications, 8, 10.1038/s41467-017-01250-8 · DOI
  56. Yofe, 2016, One library to make them all: streamlining the creation of yeast libraries via a SWAp-Tag strategy, Nature Methods, 13, 371, 10.1038/nmeth.3795 · DOI
  57. Yu, 2022, ExCel: super-resolution imaging of C. elegans with expansion microscopy, Methods in Molecular Biology, 2468, 141, 10.1007/978-1-0716-2181-3_9 · DOI
  58. Zhang, 2023, Quantitative assessment of near-infrared fluorescent proteins, Nature Methods, 20, 1605, 10.1038/s41592-023-01975-z · DOI

Metadata

Authors
Eroglu, M., & Hobert, O.
Affiliations
Howard Hughes Medical Institute (US)
Corresponding author
Hobert, Oliver
Venue
eLife
Published
2026-06-24
Volume
15
ISSN
2050-084X (ISSN-L)
OpenAlex
W7128481866
Cited by
0
References
3
Type
posted content
Language
en
Open access
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
  • Leon Levy Foundation
  • Howard Hughes Medical Institute
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

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review