title: Genetic code expansion and bioorthogonal labelling enables cell specific proteomics in an animal.
abstract: The site specific, co-translational introduction of unnatural amino acids into proteins produced in cells has been facilitated by the development of the pyrrolysyl-tRNA synthetase/tRNACUA pair. This pair can now be used to direct the site-specific incorporation of designer amino acids in E. coli, yeast, mammalian cells, and animals (the worm, C. elegans and the fly, D. melanogaster). Developments in encoding components of rapid bioorthogonal reactions are providing new opportunities for labelling and visualising proteins. A new method called stochastic orthogonal recoding of translation with chemoselective modification (SORT-M) leverages advances in genetic code expansion and bioorthogonal chemistry to label proteomes with diverse chemistry at diverse codons in E. coli, mammalian cells, and in spatially and temporally defined sets of cells in the fly. Proteomes in targeted sets of cells have been visualised by SORT-M and proteins in targeted cells have been identified by SORT-M.
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Elliott
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Ambra
authors.initials: A
authors.lastname: Bianco
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom. Electronic address: chin@mrc-lmb.cam.ac.uk.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Chin
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism; D000818:Animals; D002477:Cells / Q000378:metabolism*; D003062:Codon / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D003062:Codon; D000604:Amino Acyl-tRNA Synthetases