title: Reprogramming the genetic code in vitro.
abstract: The site-specific introduction of non-canonical amino acids into polypeptides through genetic code reprogramming has become a powerful tool for biochemical studies and bioorganic synthesis. Although a variety of such techniques have been developed, all are based on the 'mis-acylation' of tRNA molecules with non-canonical amino acids. Multiple strategies have been devised to synthesize such non-canonical aminoacyl-tRNAs; for example, those based on protein or ribozyme aminoacyl-tRNA synthetase enzymes are particularly useful. Such techniques have enabled the incorporation of hundreds of different non-canonical amino acids into polypeptides in vitro. This review discusses the development and application of in vitro genetic code reprogramming techniques, especially enzymatic mis-acylation, and examines recent efforts to engineer the translational machinery to increase the range of translatable non-canonical amino acids.
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Toby
authors.initials: T
authors.lastname: Passioura
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Bunkyo-ku, Tokyo 113-0033, Japan; Japan Science and Technology Agency (JST), Core Research for Evolutionary Science and Technology (CREST), Saitama 332-0012, Japan. Electronic address: hsuga@chem.s.u-tokyo.ac.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Suga
mesh_terms: D000818:Animals; D005815:Genetic Code; D005818:Genetic Engineering; D006801:Humans; D066298:In Vitro Techniques; D010452:Peptide Biosynthesis; D016337:RNA, Catalytic / Q000235:genetics
chemical_list: D016337:RNA, Catalytic
keywords: aminoacyl-tRNA synthetase; flexizyme; genetic code reprogramming; translation