Methods for proteome labeling
AssigneesThe Board of Trustees of the Leland Stanford Junior University, U.S. Government as represented by the Department of Veterans Affairs
InventorsAndrew YANG, Anton WYSS-CORAY, Kyle BREWER
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Abstract
Provided herein are methods for labeling the proteomes of cells, as well as methods for labeling proteins or populations of proteins produced by cells. In some embodiments, the methods comprise introducing variant aminoacyl-tRNA synthetases and noncanonical amino acids into cells. Also provided herein are polynucleotides encoding variant aminoacyl-tRNA synthetases that recognize noncanonical amino acids. The methods and compositions provided herein are useful for, among other things, identifying target cells and identifying biomarkers of interest.
Claims40
A method for labeling the proteome of a cell or a portion of the proteome of a cell, the method comprising:
- (a) introducing into the cell one or more variant aminoacyl-tRNA synthetases that recognize one or more noncanonical amino acids;
- (b) introducing one or more noncanonical amino acids into the cell;
- (c) exposing the cell to conditions such that the one or more variant aminoacyl-tRNA synthetases activate one or more tRNAs in the cell with the one or more noncanonical amino acids, thereby producing one or more noncanonical activated tRNAs, and the one or more noncanonical amino acids are integrated into the proteome by the one or more noncanonical activated tRNAs, thereby producing a modified proteome;
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- (d) contacting the modified proteome with a detectable moiety, thereby producing a labeled proteome.
A method for labeling a protein or a population of proteins produced by a cell, the method comprising:
- (a) introducing into the cell one or more variant aminoacyl-tRNA synthetases that recognize one or more noncanonical amino acids;
- (b) introducing one or more noncanonical amino acids into the cell;
- (c) exposing the cell to conditions such that the one or more variant aminoacyl-tRNA synthetases activate one or more tRNAs in the cell with the one or more noncanonical amino acids, thereby producing one or more noncanonical activated tRNAs, and the one or more noncanonical amino acids are integrated into the protein or population of proteins by the one or more noncanonical activated tRNAs, thereby producing a modified protein or a population of modified proteins;
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- (d) contacting the modified protein or population of modified proteins with a detectable moiety, thereby producing a labeled protein or a population of labeled proteins.
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An isolated polynucleotide encoding a variant aminoacyl-tRNA synthetase that recognizes a noncanonical amino acid, wherein the isolated polynucleotide comprises a nucleic acid sequence having at least about 80% identity to any one of SEQ ID NOS:1-3.
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Dependencies are inferred from the claim text.