RT Journal Article
SR Electronic
T1 FerriTag: A Genetically-Encoded Inducible Tag for Correlative Light-Electron Microscopy
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 095208
DO 10.1101/095208
A1 Clarke, Nicholas I.
A1 Royle, Stephen J.
YR 2016
UL http://biorxiv.org/content/early/2016/12/18/095208.abstract
AB 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 (CLEM). 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 CLEM by labeling proteins associated with various intracellular structures including mitochondria, plasma membrane, and clathrin-coated pits and vesicles. FerriTagging has a high signal-to-noise ratio and a labeling resolution of 10 ± 5 nm. We demonstrate how FerriTagging allows nanoscale mapping of protein location relative to a subcellular structure, and use it to detail the distribution of huntingtin-interacting protein 1 related (HIP1R) in clathrin-coated pits.