Reprogramming the genetic code in vitro

Toby Passioura; Hiroaki Suga

doi:10.1016/j.tibs.2014.07.005

Reprogramming the genetic code in vitro

Trends Biochem Sci. 2014 Sep;39(9):400-8. doi: 10.1016/j.tibs.2014.07.005. Epub 2014 Aug 13.

Authors

Toby Passioura¹, Hiroaki Suga²

Affiliations

¹ Department of Chemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Bunkyo-ku, Tokyo 113-0033, Japan.
² 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.

PMID: 25129886
DOI: 10.1016/j.tibs.2014.07.005

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.

Keywords: aminoacyl-tRNA synthetase; flexizyme; genetic code reprogramming; translation.

Publication types

Research Support, Non-U.S. Gov't
Review

MeSH terms

Animals
Genetic Code*
Genetic Engineering*
Humans
In Vitro Techniques
Peptide Biosynthesis*
RNA, Catalytic / genetics

Substances

RNA, Catalytic