ISSN: 2155-9937
Logan Brennan
Genetic Code Expansion (GCE) in combination with
orthogonal labelling provides an elegant method for labelling
proteins in live mammalian cells with organic fluorescent dyes
(Fl-dyes). A Non-Canonical Amino Acid (ncAA) with a
functional group is incorporated into the protein sequence in
response to an in-frame amber stop codon (TAG) via an
orthogonal tRNA/tRNA-synthetase pair (reviewed in Labeling is
then carried out by a rapid and specific bio orthogonal reaction
between the functional group and a Fl-dye in GCE-based
labelling. As a result, the protein of interest is fluorescently
labelled on a specific residue, eliminating the requirement for
protein tag fusions (i.e. fluorescent proteins or self-labelling
proteins) in live cell applications.