A Facile System for Encoding Unnatural Amino Acids in Mammalian Cells

Authors

  • Peng R. Chen Dr.,

    1. Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 Torry Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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    • These authors contributed equally to this work.

  • Dan Groff,

    1. Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 Torry Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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    • These authors contributed equally to this work.

  • Jiantao Guo Dr.,

    1. Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 Torry Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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  • Weijia Ou Dr.,

    1. Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121 (USA), Fax: (+1) 858-812-1746
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  • Susan Cellitti Dr.,

    1. Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121 (USA), Fax: (+1) 858-812-1746
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  • Bernhard H. Geierstanger Dr.,

    1. Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121 (USA), Fax: (+1) 858-812-1746
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  • Peter G. Schultz Dr.

    1. Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121 (USA), Fax: (+1) 858-812-1746
    2. Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 Torry Pines Road, La Jolla, CA 92037 (USA), Fax: (+1) 858-784-9440
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  • This work was supported by a grant from the U.S. National Institute of Health R01 GM062159 and the Skaggs Institute for Chemical Biology. This is manuscript 19983 of The Scripps Research Institute.

Abstract

A shuttle system has been developed to genetically encode unnatural amino acids in mammalian cells using aminoacyl-tRNA synthetases (aaRSs) evolved in E. coli. A pyrrolysyl-tRNA synthetase (PylRS) mutant was evolved in E. coli that selectively aminoacylates a cognate nonsense suppressor tRNA with a photocaged lysine derivative. Transfer of this orthogonal tRNA–aaRS pair into mammalian cells made possible the selective incorporation of this unnatural amino acid into proteins.

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