A straightforward approach for bioorthogonal labeling of proteins and organelles in live mammalian cells, using a short peptide tag.

ABSTRACT: BACKGROUND:In the high-resolution microscopy era, genetic code expansion (GCE)-based bioorthogonal labeling offers an elegant way for direct labeling of proteins in live cells with fluorescent dyes. This labeling approach is currently not broadly used in live-cell applications, partly because it needs to be adjusted to the specific protein under study. RESULTS:We present a generic, 14-residue long, N-terminal tag for GCE-based labeling of proteins in live mammalian cells. Using this tag, we generated a library of GCE-based organelle markers, demonstrating the applicability of the tag for labeling a plethora of proteins and organelles. Finally, we show that the HA epitope, used as a backbone in our tag, may be substituted with other epitopes and, in some cases, can be completely removed, re

SUBMITTER: Segal I 

PROVIDER: S-EPMC6961407 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature