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A rationally designed orthogonal synthetase for genetically encoded fluorescent amino acids.

ABSTRACT: 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.

SUBMITTER: Steinberg X

PROVIDER: S-EPMC7550906 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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A rationally designed orthogonal synthetase for genetically encoded fluorescent amino acids.

Steinberg Ximena X Galpin Jason J Nasir Gibran G Sepúlveda Romina V RV Ladron de Guevara Ernesto E Gonzalez-Nilo Fernando F Islas Leon D LD Ahern Christopher A CA Brauchi Sebastian E SE

Heliyon 20201007 10

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 ...[more]

PMID: 33083608

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