Use of eRF1 mutants in unnatural amino acid incorporation
US10738339B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Oct 21, 2015 |
| Grant date | Aug 11, 2020 |
| Priority date | — |
| Expiry date | Mar 29, 2036 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Y601/01026
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The invention relates to a method for incorporating an unnatural amino acid into a protein of interest in a eukaryotic cell, said method comprising the steps of: i) providing a eukaryotic cell expressing an orthogonal tRNA synthetase-t RNA pair, a nucleic acid sequence of interest encoding said protein of interest, and a mutant eRF1, said mutant eRF1 having amino acid sequence having at least 60% sequence identity to the human wild type eRF1 sequence of SEQ ID NO: 4, said nucleic acid sequence of interest comprising a codon recognised by the tRNA at the position for incorporation of an unnatural amino acid; ii) incubating the eukaryotic cell in the presence of an unnatural amino acid to be incorporated into a protein encoded by the nucleic acid sequence of interest, wherein said unnatural amino acid is a substrate for the orthogonal tRNA synthetase; and iii) incubating the eukaryotic cell to allow incorporation of said unnatural amino acid into the protein of interest via the orthogonal tRNA synthetase-t RNA pair. The invention also relates to uses, host cells, combinations and kits.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.