Provided are compositions comprising newly identified protein fragments of aminoacyl-tRNA synthetases, polynucleotides that encode them and complements thereof, related agents, and methods of use thereof in diagnostic, drug discovery, research, and therapeutic applications.

Provided are compositions comprising newly identified protein fragments of aminoacyl-tRNA synthetases, polynucleotides that encode them and complements thereof, related agents, and methods of use thereof in diagnostic, drug discovery, research, and therapeutic applications.

The invention provides methods and compositions for a mutein aminoacyl-tRNA synthetase that preferentially charges a tRNA with a non-natural amino acid. Also provided are methods for incorporating the non-natural amino acids, pyridinyl-amino tetrazine amino acids, into a protein, and further conjugating a biologically active adduct to the pyridinyl-amino tetrazine.

Provided are compositions comprising newly identified protein fragments of aminoacyl-tRNA synthetases, polynucleotides that encode them and complements thereof, related agents, and methods of use thereof in diagnostic, drug discovery, research, and therapeutic applications.

Provided are compositions comprising newly identified protein fragments of aminoacyl-tRNA synthetases, polynucleotides that encode them and complements thereof, related agents, and methods of use thereof in diagnostic, drug discovery, research, and therapeutic applications.

The present invention relates to a method for producing eukaryotic cell lysates for cell-free protein synthesis comprising at least one exogenous enzyme, wherein the method comprises at least the following steps: a) providing eukaryotic cells transfected with at least one donor template coding for at least one exogenous enzyme which is selected from the group comprising orthogonal aminoacyl-tRNA synthetases and viral RNA polymerases: b) cultivating the transfected cells of step a) for a predetermined period of time and subsequently harvesting the cells; and c) disrupting the harvested cells and preparing a cell lysate for cell-free protein synthesis therefrom. More specifically, the invention relates to a method for producing eukaryotic cell lysates which are capable of cell-free synthesis of a target protein comprising a non-canonical amino acid, wherein the method comprises at least the following steps: a) providing eukaryotic cells transfected with at least one donor template coding for an orthogonal aminoacyl-tRNA synthetase. i.e. an aminoacyl-tRNA synthetase which is specific for a tRNA which is not recognized by endogenous aminoacyl-tRNA synthetase in said eukaryotic cells and is specific for a corresponding non-canonical amino acid: b) cultivating the transfected cells of step a) for a predetermined period of time and subsequently harvesting the cells; and c) disrupting the harvested cells and preparing a cell lysate therefrom. Further aspects of the present invention relate in particular to eukaryotic cell lysates obtainable by the above methods as well as to a method for performing cell-free synthesis of a target protein comprising a non-canonical amino acid.

Provided herein are methods and systems for codon rewriting and replacement. In some aspects, provided herein, is a method comprising: analyzing at least a portion of a genome of an organism to identify a first plurality of codons based on at least in part on a first local context of a codon-of-interest in the genome of the organism to be rewritten; and rewriting the first plurality of codons in the genome of the organism to a second codon. Also provided herein are methods and systems for producing a synthetic genome.

Methods, compositions and kits for detecting mutated aminoacyl tRNA synthetase (aaRS) in biological samples from a subject suspect of having or suffering from a Charcot-Marie-Tooth (CMT) disease or a CMT-related disease are disclosed herein. In some embodiments, the methods include determining the amount of mutated aaRS bound to Neuropilin 1 (Nrp1). In some embodiments, methods include detection of endogenous vascular endothelial growth factor (VEGF) bound to Nrp1. Also disclosed are methods, compositions and kits for the diagnosis of a CMT or a CMT-related disease through the detection of VEGF and/or mutated aaRS in a subject.

Tetrazine non-canonical amino acids, methods for genetic encoding proteins and polypeptides using the tetrazine amino acids, proteins and polypeptides comprising the tetrazine amino acids, and compositions comprising the proteins and polypeptides having at least one post-translational modification thereof comprising in vivo reaction with the incorporated tetrazine amino acid and a second molecule.

The invention provides methods and compositions for a mutein aminoacyl-tRNA synthetase that preferentially charges a tRNA with a non-natural amino acid. Also provided are methods for incorporating the non-natural amino acids, pyridinyl-amino tetrazine amino acids, into a protein, and further conjugating a biologically active adduct to the pyridinyl-amino tetrazine.