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US 2006/0134748 A1Published application
Orthogonal suppressor tRNAs and aminoacyl-tRNA synthetases and uses thereof
OwnerUttam RajBhandary, Caroline Koehrerinventors
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OwnerUttam RajBhandary, Caroline Koehrerinventors
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The present invention provides novel orthogonal suppressor tRNAs, aminoacyl-tRNA synthetases, and suppressor tRNA/aminoacyl-tRNA synthetase pairs. In preferred embodiments the suppressor tRNAs function in mammalian cells and in certain embodiments they are not aminoacylated by any of the mammalian cytoplasmic aminoacyl-tRNA synthetases. The invention provides a novel ochre suppressor tRNA that fulfills these criteria. The invention further provides novel amber and opal suppressor tRNAs having a range of different translation efficiencies. The invention also provides mammalian cells containing the suppressor tRNAs, aminoacyl-tRNA synthetases, or both. The suppressor tRNAs may be introduced into the cell from the exterior or may be expressed by the cell. They may be aminoacylated with either a natural or an unnatural amino acid. The suppressor tRNAs, aminoacyl-tRNA synthetases, or both, may be expressed by a mammalian cell in a regulated manner. The invention further provides methods for synthesizing proteins using the inventive suppressor tRNAs and aminoacyl-tRNA synthetases and proteins produced by the methods.
An ochre suppressor tRNA that is orthogonal to a mammalian cell.
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A method for synthesizing a protein in a mammalian cell by translation of genes containing at least one stop codon within the open reading frame, the method comprising steps of:
A method for synthesizing a protein in a mammalian cell by translation of genes containing at least one stop codon within the open reading frame, the method comprising steps of:
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A kit comprising at least one suppressor tRNA that is orthogonal to a mammalian cell, or a polynucleotide or expression vector that comprises a template for synthesis of the suppressor tRNA, or both, wherein the suppressor tRNA is selected from the group consisting of:
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Dependencies are inferred from the claim text.