Steinberg, X.Galpin, J.Nasir, G.Sepulveda, R.V.Ladron de Guevara, E.Gonzalez-Nilo, F.Islas, L.D.Ahern, C.A.Brauchi, S.E.2021-07-272021-07-272020-10HeliyonOpen AccessVolume 6, Issue 10October 2020 Article number e051402405-8440http://repositorio.unab.cl/xmlui/handle/ria/19520Indexación: Scopus.The incorporation of non-canonical amino acids into proteins has emerged as a promising strategy to manipulate and study protein structure-function relationships with superior precision in vitro and in vivo. To date, fluorescent non-canonical amino acids (f-ncAA) have been successfully incorporated in proteins expressed in bacterial systems, Xenopus oocytes, and HEK-293T cells. Here, we describe the rational generation of a novel orthogonal aminoacyl-tRNA synthetase based on the E. coli tyrosine synthetase that is capable of encoding the f-ncAA tyr-coumarin in HEK-293T cells. © 2020Bioorganic Chemistry; Bioinformatics; Proteins; Biochemistry; Molecular Biology; Unnatural amino acids, aminoacyl-tRNA synthetase, coumarin, fluorescence. © 2020enAminoacyl-tRNA synthetaseBiochemistryBioinformaticsBioorganic chemistryCoumarinFluorescenceMolecular biologyProteinsUnnatural amino acidsArtículoAttribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)10.1016/j.heliyon.2020.e05140