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TitleGenetically encoded FerriTag as a specific label for cryo-electron tomography.
Journal, issue, pagesStructure, Vol. 33, Issue 12, Page 2146-22156.e4, Year 2025
Publish dateDec 4, 2025
AuthorsChang Wang / Amin Khosrozadeh / Ioan Iacovache / Benoît Zuber /
PubMed AbstractCryo-electron tomography (cryoET) provides 3D datasets of organelles and proteins at nanometer and sub-nanometer resolution. However, locating target proteins in live cells remains a significant ...Cryo-electron tomography (cryoET) provides 3D datasets of organelles and proteins at nanometer and sub-nanometer resolution. However, locating target proteins in live cells remains a significant challenge. Conventional labeling methods, such as fluorescent protein tagging and immunogold labeling, are unsuitable for small structures in vitrified samples at molecular resolution. Directly linking large, visually identifiable proteins to target proteins may alter their structure, localization, and function. To overcome this, we employed a rapamycin-induced oligomer formation system involving two tags, FK506 binding protein (FKBP) and FKBP-rapamycin binding (FRB), which bind in the presence of rapamycin. FKBP is linked to the target protein, while FRB is linked to ferritin, a large (10-12 nm) iron-binding complex that creates strong contrast in cryoET. Upon adding rapamycin to the cell medium, the iron-loaded ferritin accurately marks the target protein location. As in situ cryoET with subtomogram averaging advances, our method addresses the persistent challenge of locating target proteins in live cells.
External linksStructure / PubMed:40925364
MethodsEM (tomography)
Structure data

EMDB-54356: Genetically Encoded FerriTag as a Specific Label for Cryo-Electron Tomography
Method: EM (tomography)

Source
  • Homo sapiens (human)

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