10.1021/acsnano.8b06047.s001
Kim-A. Saal
Kim-A.
Saal
Frank Richter
Frank
Richter
Peter Rehling
Peter
Rehling
Silvio O. Rizzoli
Silvio O.
Rizzoli
Combined
Use of Unnatural Amino Acids Enables Dual-Color
Super-Resolution Imaging of Proteins <i>via</i> Click Chemistry
American Chemical Society
2018
fluorophore conjugation
Click Chemistry
cell sets
protein
dual-color super-resolution microscopy
optimized UAAs
POI
click chemistry
Unnatural Amino Acids Enables Dual-Color Super-Resolution Imaging
click-based imaging
Such requirements
Amber codon
Fuse 2Click
imaging resolution
2018-12-10 00:00:00
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Combined_Use_of_Unnatural_Amino_Acids_Enables_Dual-Color_Super-Resolution_Imaging_of_Proteins_i_via_i_Click_Chemistry/7464575
Recent
advances in optical nanoscopy have brought the imaging resolution
to the size of the individual macromolecules, thereby setting stringent
requirements for the fluorescent labels. Such requirements are optimally
fulfilled by the incorporation of unnatural amino acids (UAAs) in
the proteins of interest (POIs), followed by fluorophore conjugation <i>via</i> click chemistry. However, this approach has been limited
to single POIs in mammalian cells. Here we solve this problem by incorporating
different UAAs in different POIs, which are expressed in independent
cell sets. The cells are then fused, thereby combining the different
proteins and organelles, and are easily imaged by dual-color super-resolution
microscopy. This procedure, which we termed Fuse2Click, is simple,
requires only the well-established Amber codon, and allows the use
of all previously optimized UAAs and tRNA/RS pairs. This should render
it a tool of choice for multicolor click-based imaging.