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- Title
Genetic incorporation of unnatural amino acids into proteins in mammalian cells.
- Authors
Wenshe Liu; Brock, Ansgar; Shuo Chen; Shuibing Chen; Schultz, Peter G.
- Abstract
We developed a general approach that allows unnatural amino acids with diverse physicochemical and biological properties to be genetically encoded in mammalian cells. A mutant Escherichia coli aminoacyl-tRNA synthetase (aaRS) is first evolved in yeast to selectively aminoacylate its tRNA with the unnatural amino acid of interest. This mutant aaRS together with an amber suppressor tRNA from Bacillus stearothermophilus is then used to site-specifically incorporate the unnatural amino acid into a protein in mammalian cells in response to an amber nonsense codon. We independently incorporated six unnatural amino acids into GFP expressed in CHO cells with efficiencies up to 1 μg protein per 2 × 107 cells; mass spectrometry confirmed a high translational fidelity for the unnatural amino acid. This methodology should facilitate the introduction of biological probes into proteins for cellular studies and may ultimately facilitate the synthesis of therapeutic proteins containing unnatural amino acids in mammalian cells.
- Publication
Nature Methods, 2007, Vol 4, Issue 3, p239
- ISSN
1548-7091
- Publication type
Academic Journal
- DOI
10.1038/nmeth1016