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Abstrakt

Molecular Tracking and Localization of Bio Orthogonally Labeled Plasma Membrane Proteins in Live Cells

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.