Using unnatural amino acids to selectively label proteins for cellular imaging: a cell biologist viewpoint

Research output: Contribution to journalReview articlepeer-review

12 Scopus citations

Abstract

Twenty-five years ago, GFP revolutionized the field of cell biology by enabling scientists to visualize, for the first time, proteins in living cells. However, when it comes to current, state-of-the-art imaging technologies, fluorescent proteins (such as GFP) have several limitations that result from their size and photophysics. Over the past decade, an elegant, alternative approach, which is based on the direct labeling of proteins with fluorescent dyes and is compatible with live-cell and super-resolution imaging applications, has been introduced. In this approach, an unnatural amino acid that can covalently bind a fluorescent dye is incorporated into the coding sequence of a protein. The protein of interest is thereby site-specifically fluorescently labeled inside the cell, eliminating the need for protein- or peptide-labeling tags. Whether this labeling approach will change cell biology research is currently unclear, but it clearly has the potential to do so. In this short review, a general overview of this approach is provided, focusing on the imaging of site-specifically labeled proteins in mammalian tissue culture cells, and highlighting its advantages and limitations for cellular imaging.

Original languageEnglish
Pages (from-to)1107-1117
Number of pages11
JournalFEBS Journal
Volume288
Issue number4
DOIs
StatePublished - 1 Feb 2021

Keywords

  • bioorthogonal reactions
  • click chemistry
  • fluorescent dyes
  • genetic code expansion
  • light microscopy
  • noncanonical amino acids
  • protein labeling

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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