Genetic code expansion with unnatural amino acids (UAAs) has significantly broadened the chemical repertoire of proteins. Applications of this method in mammalian cells include probing of molecular interactions, conditional control of biological processes, and new strategies for therapeutics and vaccines. A number of methods have been developed for transient UAA mutagenesis in mammalian cells, each with unique features and advantages. All have in common a need to deliver genes encoding additional protein biosynthetic machinery (an orthogonal tRNA/tRNA synthetase pair) and a gene for the protein of interest. In this study, we present a comparative evaluation of select plasmid-based genetic code expansion systems and a detailed analysis of suppression efficiency with different UAAs and in different cell lines.

非天然氨基酸 (UAA) 的遗传密码扩展显着拓宽了蛋白质的化学库。这种方法在哺乳动物细胞中的应用包括分子相互作用的探测、生物过程的条件控制以及治疗和疫苗的新策略。已经开发了多种用于哺乳动物细胞中瞬时 UAA 诱变的方法，每种方法都具有独特的特征和优势。所有人都需要提供编码额外蛋白质生物合成机制（正交 tRNA/tRNA 合成酶对）的基因和目标蛋白质的基因。在这项研究中，我们对选择的基于质粒的遗传密码扩展系统进行了比较评估，并对不同 UAA 和不同细胞系中的抑制效率进行了详细分析。