ORTHOGONAL SUPPRESSOR TRNAS AND AMINOACYL-TRNA SYNTHETASES AND USES THEREOF

摘要

The present invention provides novel orthogonal suppressor tRNAs, aminoacyl­tRNA 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 aminoacyl­tRNA 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 efficiences. 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.