Institut Pasteur

The Scripps Research Institute

Evologic Gmbh

Volker Doring

Philippe Marliere

Paul R Schimmel

Tamara L Hendrickson

Henning Mootz

Valerie De Crecy-Lagard

Leslie A Nangle

The present invention relates to a method for providing bacterial or yeast cells with the capacity to produce a protein, the amino acid sequence of which comprises at least one unconventional amino acid. The method involves (a) introducing at least one missense mutation in a target codon of a gene encoding a protein required for the growth of the bacterial or yeast cells, where the mutated protein synthesized from the mutated gene is not functional in the bacterial or yeast cells. The method also involves (b) selecting the bacterial or yeast cells obtained in (a) in a culture medium which (1) does not contain a nutrient compensating for the loss of functionality of the mutated protein and (2) contains an unconventional amino acid which restores the functionality of the protein required for growth of the bacterial or yeast cells, in which the unconventional amino acid is that encoded by the target codon. The method also involves culturing the bacterial or yeast cells obtained in (b) in a culture medium containing the amino acid encoded by the target codon.

Cells and method for producing proteins comprising an unconventional amino acid

The present invention is directed to a method to diversify the chemical composition of proteins produced in vivo comprising the step of disabling, particularly by mutagenesis, the editing function of one of its aminoacyl tRNA synthetases. The present invention is also directed to nucleic acid sequences encoding such mutated aminoacyl tRNA synthetases having their editing site mutated and capable of mischarging its cognate tRNA with a noncanonical amino acid. Also described herein is an improved method for obtaining transformed cells capable of synthetizing in vivo proteins comprising at least a noncanonical amino acid and their use for the production of such proteins.

Method for diversifying the chemical composition of proteins produced in vivo by genetically disabling the editing function of their aminoacyl tRNA synthetases

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Paris, France

Volker Doring