Abstract
A current challenge is to develop tags to precisely visualize proteins in cells by light and electron microscopy. Here, we introduce FerriTag, a genetically-encoded chemically-inducible tag for correlative light-electron microscopy. FerriTag is a fluorescent recombinant electron-dense ferritin particle that can be attached to a protein-of-interest using rapamycin-induced heterodimerization. We demonstrate the utility of FerriTag for correlative light-electron microscopy by labeling proteins associated with various intracellular structures including mitochondria, plasma membrane, and clathrin-coated pits and vesicles. FerriTagging has a good signal-to-noise ratio and a labeling resolution of approximately 10 nm. We demonstrate how FerriTagging allows nanoscale mapping of protein location relative to a subcellular structure, and use it to detail the distribution and conformation of huntingtin-interacting protein 1 related (HIP1R) in and around clathrin-coated pits.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Cell Membrane / metabolism
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Cell Membrane / ultrastructure
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Clathrin-Coated Vesicles / metabolism
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Clathrin-Coated Vesicles / ultrastructure
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Coated Pits, Cell-Membrane / metabolism
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Coated Pits, Cell-Membrane / ultrastructure
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Ferritins / chemistry
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Ferritins / genetics*
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Ferritins / metabolism
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Fluorescent Dyes / chemistry*
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Gene Expression
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HeLa Cells
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Humans
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Microfilament Proteins
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Microscopy, Electron / methods*
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Mitochondria / metabolism
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Mitochondria / ultrastructure
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Protein Multimerization
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Signal-To-Noise Ratio
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Sirolimus / chemistry*
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Staining and Labeling / methods*
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Vesicular Transport Proteins / analysis
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Vesicular Transport Proteins / genetics
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Vesicular Transport Proteins / metabolism
Substances
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Adaptor Proteins, Signal Transducing
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Fluorescent Dyes
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HIP1R protein, human
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Microfilament Proteins
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Recombinant Proteins
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Vesicular Transport Proteins
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Ferritins
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Sirolimus