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Development of mammalian cell logic gates controlled by unnatural amino acids

Mills, Emily M., Barlow, Victoria L., Jones, Arwyn T. ORCID: https://orcid.org/0000-0003-2781-8905 and Tsai, Yu-Hsuan ORCID: https://orcid.org/0000-0003-0589-5088 2021. Development of mammalian cell logic gates controlled by unnatural amino acids. Cell Reports Methods 1 (6) , 100073. 10.1016/j.crmeth.2021.100073

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Abstract

Genetic code expansion employing an orthogonal aminoacyl-tRNA synthetase/tRNA pair for site-specific unnatural amino acid incorporation has a wide range of applications. However, a blank (orthogonal) codon is needed for each orthogonal tRNA. Although the amber stop codon is commonly used as the blank codon, the use of stop codons as the blank codons inevitably limits the number of possible different unnatural amino acids to be incorporated into proteins in a single cell. To overcome this limitation, we evaluated the capability of 11 Pyl tRNA variants for decoding quadruplet codons in mammalian cells. With efficient quadruplet-decoding orthogonal tRNA variants available, it is possible for simultaneous incorporation of more than three different unnatural amino acids in mammalian cells.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Chemistry
Pharmacy
Additional Information: This is an open access article under the terms of the CC-BY Attribution 4.0 International license.
Publisher: Elsevier
ISSN: 2667-2375
Funders: BBSRC, EPSRC
Date of First Compliant Deposit: 30 September 2021
Date of Acceptance: 13 August 2021
Last Modified: 09 Jul 2023 17:33
URI: https://orca.cardiff.ac.uk/id/eprint/144571

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