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Cited by 45RNA Research and SplicingProtein Structure and DynamicsAdvanced Fluorescence Microscopy Techniques

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Fluorescent protein and peptide tags alter condensate formation and dynamics in vivo and in vitro

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Dörner, Kerstin, Gut, Michelle Jennifer, Overwijn, Daan, Cao, Fan, Siketanc, Matej, Heinrich, Stephanie, Beuret, Nicole, Meyer, Justin, Sharpe, Timothy, Lindorff-Larsen, Kresten, Hondele, Maria·2026·EMBO Reports
DOI 10.1038/s44319-025-00626-yPMC PMC12796366

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Abstract

Fluorescent proteins and peptide tags are essential tools in cellular biology, but can alter the biochemical and physiological behavior of target proteins. Biomolecular condensates, which have emerged as key elements of cellular organization, are suggested to provide robustness to cells, yet they can also respond sensitively to small changes in environmental conditions including tags. Here, we investigate the effects of over twenty widely used tags on condensate formation in vitro, in cells, in various model organisms and by computational modelling. We find that tagging strongly influences condensation for some proteins, while others remain unaffected. Effects vary, with some tags enhancing and others decreasing condensation, with the outcome depending on the protein being tagged. Coarse-grained simulations suggest that the charge of the fluorescent protein tags is a critical factor modulating condensation behavior. Together, our results underscore the need to tag with caution and highlight the importance of careful experimental design and interpretation, especially in condensate studies, but also suggest that fluorescent protein tags could serve as a tool to modulate condensate properties.

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Metadata

Authors
Dörner, K., Gut, M. J., Overwijn, D., Cao, F., Siketanc, M., Heinrich, S., Beuret, N., Meyer, J., Sharpe, T., Lindorff-Larsen, K., & Hondele, M.
Affiliations
  • University of Basel (CH)
  • University of Copenhagen (DK)
Corresponding author
Hondele, Maria
ORCID
Venue
EMBO Reports
Publisher
Springer Science and Business Media LLC
Published
2025-11-20
Volume
27
Issue
1
Pages
89-121
ISSN
  • 1469-221X (ISSN-L)
  • 1469-3178
DOI
10.1038/s44319-025-00626-y
PMC
PMC12796366
OpenAlex
W4416416816
Cited by
45
References
1
Citations by year
2026: 27 · 2025: 17
FWCI
13.23
Type
journal article
Language
en
Open access
Licence
Locations
  • EMBO Reports (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • EC | European Research Council (ERC-ST2020 950262)
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (PCEFP3_187052)
  • Novo Nordisk Fonden (NNF18OC0033950)
  • China Scholarship Council (CSC 202206340019)
  • Universität Basel (Fellowship for Excellence, Fellowship for Excellece)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
Keywords
biomolecular condensates, fluorescent protein tags, peptide tags, condensate formation, protein charge, coarse-grained simulation, model organisms, cell organization
MeSH terms
Biomolecular Condensates, Animals, Humans, Luminescent Proteins, Peptides

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