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Related Experiment Video

Updated: Oct 9, 2025

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A proximity labeling protocol to probe proximity interactions in C. elegans.

Ariana D Sanchez1, Jessica L Feldman1

  • 1Department of Biology, Stanford University, Stanford, CA 94305, USA.

STAR Protocols
|December 20, 2021
PubMed
Summary

This study presents a new method for proximity labeling (PL) in living organisms. The protocol enables targeted protein network profiling in specific tissues of Caenorhabditis elegans.

Keywords:
Cell BiologyDevelopmental biologyMicroscopyModel OrganismsMolecular BiologyMolecular/Chemical ProbesProtein BiochemistryProteomics

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Enzyme-catalyzed proximity labeling (PL) is crucial for identifying protein-protein interactions within cells.
  • Historically, PL techniques faced limitations for application in living multicellular organisms.
  • Technical challenges hindered the widespread use of PL in complex living systems.

Purpose of the Study:

  • To develop a practical protocol for proximity labeling in living Caenorhabditis elegans.
  • To adapt PL techniques for use in multicellular organisms.
  • To enable targeted protein network profiling in vivo.

Main Methods:

  • Utilized the biotin ligase mutant enzyme TurboID for proximity labeling.
  • Applied the protocol to living C. elegans.
  • Focused on non-centrosomal Microtubule Organizing Centers (ncMTOCs) in intestinal cells for tissue-specific labeling.

Main Results:

  • Successfully demonstrated proximity labeling in a tissue-specific and region-specific manner within C. elegans.
  • Showcased the feasibility of applying PL to non-centrosomal MTOCs in intestinal cells.
  • Validated the protocol for targeted in vivo protein network analysis.

Conclusions:

  • The developed protocol facilitates proximity labeling in living C. elegans.
  • This method allows for targeted in vivo protein network profiling.
  • The protocol is a valuable tool for studying protein interactions in multicellular organisms.