Expanding and reprogramming the genetic code of cells and animals

• Published in: Annual review of biochemistry Vol. 83; p. 379
• Main Author: Chin, Jason W
• Format: Journal Article
• Language: English
• Published: United States 01.01.2014
• Subjects: Amino Acids - chemistry; Amino Acyl-tRNA Synthetases - chemistry; Animals; Caenorhabditis elegans; Drosophila melanogaster; Escherichia coli - genetics; Evolution, Molecular; Gene Deletion; Genetic Code; Genome; Protein Engineering - methods; Ribosomes - chemistry; RNA, Transfer - chemistry; Saccharomyces cerevisiae - genetics; synthetic biology; protein labeling; bioorthogonal reaction; tRNA; amino acid; aminoacyl-tRNA synthetase; posttranslational modification; protein chemistry; ribosome; genetic code expansion
• ISSN: 1545-4509
• DOI: 10.1146/annurev-biochem-060713-035737

Genetic code expansion and reprogramming enable the site-specific incorporation of diverse designer amino acids into proteins produced in cells and animals. Recent advances are enhancing the efficiency of unnatural amino acid incorporation by creating and evolving orthogonal ribosomes and manipulating the genome. Increasing the number of distinct amino acids that can be site-specifically encoded has been facilitated by the evolution of orthogonal quadruplet decoding ribosomes and the discovery of mutually orthogonal synthetase/tRNA pairs. Rapid progress in moving genetic code expansion from bacteria to eukaryotic cells and animals (C. elegans and D. melanogaster) and the incorporation of useful unnatural amino acids has been aided by the development and application of the pyrrolysyl-transfer RNA (tRNA) synthetase/tRNA pair for unnatural amino acid incorporation. Combining chemoselective reactions with encoded amino acids has facilitated the installation of posttranslational modifications, as well as rapid derivatization with diverse fluorophores for imaging.