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Related Experiment Video

Updated: Oct 26, 2025

Fluorescence Labeling to Visualize Low-Expressed Proteins in Zebrafish
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A Toolbox for Efficient Proximity-Dependent Biotinylation in Zebrafish Embryos.

Shimon M Rosenthal1, Tvisha Misra2, Hala Abdouni3

  • 1Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada; Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, Toronto, Ontario, Canada; Lunenfeld-Tanenbaum Research Institute at Mount Sinai Hospital, Sinai Health, Toronto, Ontario, Canada.

Molecular & Cellular Proteomics : MCP
|July 31, 2021
PubMed
Summary

Researchers developed in vivo proximity labeling in zebrafish, a vertebrate model, to discover protein organization and function. This new proteomics tool aids in understanding cellular structures and developmental processes.

Keywords:
BioIDProximity-dependent biotinylationTurboIDminiTurbozebrafish embryo

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Area of Science:

  • Proteomics
  • Molecular Biology
  • Developmental Biology

Background:

  • Understanding protein organization is crucial for elucidating protein function.
  • Proximity-dependent labeling methods like BioID have advanced protein discovery in cell culture.
  • Limited studies exist on in vivo proximity labeling in living vertebrates.

Purpose of the Study:

  • To adapt and optimize in vivo proximity labeling techniques for protein discovery in zebrafish.
  • To establish protocols for TurboID and miniTurbo labeling in early zebrafish embryos.
  • To enable time-resolved analysis of protein interactions during development.

Main Methods:

  • Utilized lamin A (LMNA) as bait and green fluorescent protein (GFP) as a negative control.
  • Developed both mRNA injection and heat shock-inducible transgenic systems for labeling.
  • Provided biotin directly in egg water, optimizing labeling time to 12 hours.

Main Results:

  • Successfully implemented and benchmarked in vivo TurboID and miniTurbo labeling in zebrafish embryos.
  • Identified proximal partners of LMNA, enriched for nuclear envelope and nuclear membrane proteins.
  • Found orthologs of human lamin A proximity partners, validating the approach.

Conclusions:

  • The developed tools and protocols enable powerful proteomics approaches in zebrafish.
  • This method complements genetic tools for studying protein organization and function in vivo.
  • Facilitates comparative proteomic studies between zebrafish and mammalian systems.