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PreprintCited by 5Advanced Fluorescence Microscopy TechniquesMolecular Sensors and Ion DetectionLuminescence and Fluorescent Materials

The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.

A tripartite chemogenetic fluorescent reporter: design and evaluation for visualizing ternary protein complexes

Open access
Bottone, Sara, Broch, Fanny, Gedeon, Antoine, Brion, Aurélien, El Hajji, Lina, Benaissa, Hela, Gautier, Arnaud·2023·Nature Communications
DOI 10.1101/2023.10.19.563144PMC PMC12307597

Preprint — published as 10.1038/s41467-025-62241-8

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Abstract

Most cellular processes are carried out by multiprotein assemblies. Although various molecular tools exist to visualize binary protein interactions in live cells, the visualization of multiprotein complexes remains a challenge. Here, we report the engineering of a complementation-based approach allowing one to visualize the interaction of three proteins through effective proximity-induced complementation of three fragments of pFAST, a chemogenetic fluorescent reporter that binds and stabilizes the fluorescent state of fluorogenic chromophores (so-called fluorogens). This tripartite-split-pFAST allowed the observation of dynamic ternary protein complexes in the cytosol, at the plasma membrane, in the nucleus and at the junction of multiple organelles, opening great prospects to study the role and function of multiprotein complexes in live cells and in various biologically relevant contexts.

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Metadata

Authors
Bottone, S., Broch, F., Gedeon, A., Brion, A., El Hajji, L., Benaissa, H., & Gautier, A.
Affiliations
Centre National de la Recherche Scientifique (FR)
Corresponding author
Gautier, Arnaud
ORCID
Venue
Nature Communications
Publisher
openRxiv
Published
2023-10-20
Volume
16
Issue
1
Pages
6955-6955
ISSN
2041-1723 (ISSN-L)
DOI
10.1101/2023.10.19.563144
PMC
PMC12307597
OpenAlex
W4412785354
Cited by
5
References
1
Citations by year
2026: 3 · 2025: 2
FWCI
1.88
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • HAL (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Institut Universitaire de France
  • Sorbonne Université
  • H2020 European Research Council (724705)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Physical Sciences › Chemistry › Spectroscopy › Molecular Sensors and Ion Detection
  • Physical Sciences › Materials Science › Materials Chemistry › Luminescence and Fluorescent Materials
Keywords
ternary protein complex, protein complexes, fluorogens, split fluorescent proteins, live-cell imaging, protein-protein interaction, plasma membrane
MeSH terms
Chemogenetics, Cell Membrane, Cell Nucleus, Cytosol, Fluorescent Dyes, HeLa Cells, Humans, Microscopy, Fluorescence, Genes, Reporter, Multiprotein Complexes, HEK293 Cells

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