Hybrid suppressor tRNA for vertebrate cells
First Claim
1. An isolated vertebrate cell or cell line comprising a polynucleotide comprising the nucleotide sequence as set forth in SEQ ID NO:
- 88, wherein said nucleotide sequence encodes an orthogonal tRNA.
2 Assignments
0 Petitions
Accused Products
Abstract
This invention provides compositions and methods for producing translational components that expand the number of genetically encoded amino acids in vertebrate cells. The components include orthogonal tRNA'"'"'s, orthogonal aminoacyl-tRNA synthetases, orthogonal pairs of tRNA'"'"'s/synthetases and unnatural amino acids. Proteins and methods of producing proteins with unnatural amino acids in vertebrate cells are also provided. The present invention provides vertebrate cells with translation components, e.g., pairs of orthogonal aminoacyl-tRNA synthetases (O-RSs) and orthogonal tRNA'"'"'s (O-tRNA'"'"'s) and individual components thereof, that are used in vertebrate protein biosynthetic machinery to incorporate an unnatural amino acid in a growing polypeptide chain, in a vertebrate cell.
153 Citations
26 Claims
-
1. An isolated vertebrate cell or cell line comprising a polynucleotide comprising the nucleotide sequence as set forth in SEQ ID NO:
- 88, wherein said nucleotide sequence encodes an orthogonal tRNA.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 21, 22, 23)
-
20. An isolated A vertebrate cell comprising an orthogonal tRNA encoded by SEQ ID NO:
- 88, an orthogonal aminoacyl tRNA synthetase (O-RS), a non-natural amino acid, and a nucleic acid that comprises a polynucleotide that encodes a polypeptide of interest, wherein the polynucleotide comprises a selector codon that is recognized by the tRNA, wherein the O-RS preferentially aminoacylates the tRNA with the non-natural amino acid in the vertebrate cell, and wherein the cell produces the polypeptide of interest in the presence of the non-natural amino acid.
-
24. A kit for producing a protein that comprises at least one unnatural amino acid, the kit comprising:
- a container containing an orthogonal tRNA encoded by the polynucleotide sequence set forth in SEQ ID NO;
88 and wherein the orthogonal tRNA mediates the incorporation of the unnatural amino acid into an expressed protein. - View Dependent Claims (25, 26)
- a container containing an orthogonal tRNA encoded by the polynucleotide sequence set forth in SEQ ID NO;
Specification