Adding New Chemistries to the Genetic Code

Liu, Chang C.; Schultz, Peter G.

doi:10.1146/annurev.biochem.052308.105824

Cited by 1,582 publications

(1,563 citation statements)

References 189 publications

(203 reference statements)

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“…The UAAs trans-cyclooctene lysine (TCO) and simple cyclooctyne lysine (SCO) (Fig. 6) were incorporated by amber (TAG) stop-codon suppression according to the expanded genetic code concept 38 . We used the previously reported tRNA Pyl /PylRS AF pair and GFP 39TAG (TAG codon in position 39) in our study to express GFP TAG UAA and so genetically encode the respective UAAs 36,39 .…”

Section: Resultsmentioning

confidence: 99%

A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins

et al. 2013

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The ideal fluorescent probe for bioimaging is bright, absorbs at long wavelengths and can be implemented flexibly in living cells and in vivo. However, the design of synthetic fluorophores that combine all of these properties has proved to be extremely difficult. Here, we introduce a biocompatible near-infrared silicon-rhodamine probe that can be coupled specifically to proteins using different labelling techniques. Importantly, its high permeability and fluorogenic character permit the imaging of proteins in living cells and tissues, and its brightness and photostability make it ideally suited for live-cell super-resolution microscopy. The excellent spectroscopic properties of the probe combined with its ease of use in live-cell applications make it a powerful new tool for bioimaging.

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Section: Resultsmentioning

confidence: 99%

A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins

et al. 2013

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“…The enrichment of clones harboring the TAG codon during a selection should indicate that an unnatural amino acid confers a selective advantage. This library was cotransformed into E. coli BL21(DE3) along with the pEVOL MjRS∕MjtRNA Tyr CUA expression system which encodes an evolved orthogonal aminoacyl-tRNA synthetase (aaRS) and a cognate amber suppressor tRNA CUA (37) that incorporates a desired unnatural amino acid site-specifically in response to the amber codon (38). A selection was then carried out for cyclic peptides that inhibit HIV protease with p-benzoylphenylalanine (pBzF) as a 21st amino acid.…”

Section: Resultsmentioning

confidence: 99%

Evolution of cyclic peptide protease inhibitors

Young

Ahmad

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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127

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We report a bacterial system for the evolution of cyclic peptides that makes use of an expanded set of amino acid building blocks. Orthogonal aminoacyl-tRNA synthetase/tRNA CUA pairs, together with a split intein system were used to biosynthesize a library of ribosomal peptides containing amino acids with unique structures and reactivities. This peptide library was subsequently used to evolve an inhibitor of HIV protease using a selection based on cellular viability. Two of three cyclic peptides isolated after two rounds of selection contained the keto amino acid p -benzoylphenylalanine ( p BzF). The most potent peptide (G12: GIXVSL; X = p BzF) inhibited HIV protease through the formation of a covalent Schiff base adduct of the p BzF residue with the ϵ-amino group of Lys 14 on the protease. This result suggests that an expanded genetic code can confer an evolutionary advantage in response to selective pressure. Moreover, the combination of natural evolutionary processes with chemically biased building blocks provides another strategy for the generation of biologically active peptides using microbial systems.

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“…Two polyspecific tRNA/aminoacyl-tRNA synthetase pairs were inserted into this expression system, enabling the site-specific incorporation of a variety of unnatural amino acids with novel chemical and biological properties into proteins. into proteins allows the introduction of biochemical or biophysical probes at defined sites in the proteomes of living cells (1). The UAA of interest is genetically encoded by a reassigned nonsense (typically TAG) or frameshift codon, and is cotranslationally incorporated into proteins using an orthogonal UAAspecific tRNA/aminoacyl-tRNA synthetase (tRNA/aaRS) pair.…”

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confidence: 99%

Efficient viral delivery system for unnatural amino acid mutagenesis in mammalian cells

Chatterjee

Xiao

Bollong

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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109

132

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Here we report the development of a baculovirus-based delivery system that enables the efficient incorporation of unnatural amino acids into proteins in mammalian cells. We have exploited the large cargo-capacity (>30 kb) and stability of the double-stranded DNA genome of baculovirus to deliver to a variety of cell types all of the components required to genetically incorporate novel amino acids. These include the engineered tRNA/aminoacyl-tRNA synthetase pair and the nonsense mutant of the target gene. Mammalian cell transduction efficiency of baculovirus was significantly improved by incorporating genetic elements from mammalian viruses. Two polyspecific tRNA/aminoacyl-tRNA synthetase pairs were inserted into this expression system, enabling the site-specific incorporation of a variety of unnatural amino acids with novel chemical and biological properties into proteins.genetic code | viral vector | nonstandard amino acid | orthogonal

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Adding New Chemistries to the Genetic Code

Cited by 1,582 publications

References 189 publications

A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins

A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins

Evolution of cyclic peptide protease inhibitors

Efficient viral delivery system for unnatural amino acid mutagenesis in mammalian cells

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