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bioRxiv · 10.1101/065227

A Rationally Designed Aminoacyl-tRNA Synthetase for Genetically Encoded Fluorescent Amino Acids

Ximena Steinberg·Jason Galpin·Gibran Nasir·Jose Sepulveda-Ugarte·Romina Sepulveda·Fernando Gonzalez-Nilo·Leon D Islas·Christopher A Ahern·Sebastian E Brauchi
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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 an orthogonal aminoacyltRNA synthetase based on the E. coli tyrosine synthetase that is capable of encoding the f-ncAA tyr-coumarin in HEK-293T cells.

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Ximena Steinberg, Jason Galpin, Gibran Nasir, Jose Sepulveda-Ugarte, Romina Sepulveda, Fernando Gonzalez-Nilo, Leon D Islas, Christopher A Ahern, Sebastian E Brauchi. 2016-07-22. A Rationally Designed Aminoacyl-tRNA Synthetase for Genetically Encoded Fluorescent Amino Acids. https://doi.org/10.1101/065227

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