Rare codon recoding for efficient noncanonical amino acid incorporation in mammalian cells

• 成果类型: Article
• 署名作者: Ding, Wenlong; Yu, Wei; Chen, Yulin; Lao, Lihui; Fang, Yu; Fang, Chengzhu; Zhao, Hongxia; Yang, Bo; Lin, Shixian
• 署名单位: Zhejiang University; Zhejiang University; Zhejiang University; Zhejiang University; Zhejiang University
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-8537
• DOI: 10.1126/science.adm8143
• 发表日期: 2024-06-07
• 页码: 1134-1142
• 关键词: site-specific incorporation; suppressor transfer-rnas; genetic-code; directed evolution; expansion; activation; proteins; platform; lysine

摘要

The ability to genetically encode noncanonical amino acids (ncAAs) has empowered proteins with improved or previously unknown properties. However, existing strategies in mammalian cells rely on the introduction of a blank codon to incorporate ncAAs, which is inefficient and limits their widespread applications. In this study, we developed a rare codon recoding strategy that takes advantage of the relative rarity of the TCG codon to achieve highly selective and efficient ncAA incorporation through systematic engineering and big data-model predictions. We highlight the broad utility of this strategy for the incorporation of dozens of ncAAs into various functional proteins at the wild-type protein expression levels, as well as the synthesis of proteins with up to six-site ncAAs or four distinct ncAAs in mammalian cells for downstream applications.