Review Article | Open Access

Volume 2022 |Article ID 9896125 | https://doi.org/10.34133/2022/9896125

Improving the Efficiency and Orthogonality of Genetic Code Expansion

Xian Fu iD ,¹ Yijian Huang,^1,3 Yue Shen iD ^1,2,4

¹BGI-Shenzhen, Shenzhen 518083, China
²Guangdong Provincial Key Laboratory of Genome Read and Write, Shenzhen 518120, China
³School of Future Technology, University of Chinese Academy of Sciences, Beijing 100049, China
⁴Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China

Abstract

The site-specific incorporation of the noncanonical amino acid (ncAA) into proteins via genetic code expansion (GCE) has enabled the development of new and powerful ways to learn, regulate, and evolve biological functions in vivo. However, cellular biosynthesis of ncAA-containing proteins with high efficiency and fidelity is a formidable challenge. In this review, we summarize up-to-date progress towards improving the efficiency and orthogonality of GCE and enhancing intracellular compatibility of introduced translation machinery in the living cells by creation and optimization of orthogonal translation components, constructing genomically recoded organism (GRO), utilization of unnatural base pairs (UBP) and quadruplet codons (four-base codons), and spatial separation of orthogonal translation.