Chimeric design of pyrrolysyl-tRNA synthetase/tRNA pairs and canonical synthetase/tRNA pairs for genetic code expansion
Chimeric design of pyrrolysyl-tRNA synthetase/tRNA pairs and canonical synthetase/tRNA pairs for genetic code expansion
复制标题
用于遗传密码扩展的吡咯赖氨酰-tRNA合成酶/tRNA对和典型合成酶/tRNA对的嵌合设计
DOI：
10.1038/s41467-020-16898-y
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发表时间：
2020-06-22
影响因子：
16.6
通讯作者：
Lin, Shixian
中科院分区：
文献类型：
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作者：
Ding, Wenlong;Zhao, Hongxia;Lin, Shixian
正交氨基酰基-TRNA合成酶/tRNA对是非自然氨基酸特异性掺入的至关重要的先决条件。由于其高密码子抑制效率和完全正交性，吡咯基-TRNA合成酶/吡咯基-TRNA对是当前真核生物和原核生物遗传代码扩展的理想系统。迫切需要发现或设计其他完全正交翻译系统。在这里，通过从吡咯氨氨酸系统中移植关键的正交成分的理性嵌合体设计，我们创建了多个嵌合tRNA合成酶/嵌合tRNA对，包括嵌合内组氨酸，苯丙氨酸和丙氨酸系统。我们进一步表明，这些工程化的嵌合系统是正交和高效的，具有与吡咯氨氨酸系统相当的灵活性。此外，嵌合体苯丙氨酸系统可以在大肠杆菌和哺乳动物细胞中有效地融合一组苯丙氨酸，酪氨酸和色氨酸类似物。这些芳族氨基酸类似，具有独特的特性和特性，包括荧光，翻译后修饰。正交氨基酰基-TRNA合成酶/tRNA对对于以特定地点的方式掺入非天然氨基酸至关重要。在这里，作者使用有理嵌合体设计来创建多个有效的对，这些对在细菌和哺乳动物系统中起作用，以扩展遗传代码。
An orthogonal aminoacyl-tRNA synthetase/tRNA pair is a crucial prerequisite for site-specific incorporation of unnatural amino acids. Due to its high codon suppression efficiency and full orthogonality, the pyrrolysyl-tRNA synthetase/pyrrolysyl-tRNA pair is currently the ideal system for genetic code expansion in both eukaryotes and prokaryotes. There is a pressing need to discover or engineer other fully orthogonal translation systems. Here, through rational chimera design by transplanting the key orthogonal components from the pyrrolysine system, we create multiple chimeric tRNA synthetase/chimeric tRNA pairs, including chimera histidine, phenylalanine, and alanine systems. We further show that these engineered chimeric systems are orthogonal and highly efficient with comparable flexibility to the pyrrolysine system. Besides, the chimera phenylalanine system can incorporate a group of phenylalanine, tyrosine, and tryptophan analogues efficiently in both E. coli and mammalian cells. These aromatic amino acids analogous exhibit unique properties and characteristics, including fluorescence, post-translation modification. Orthogonal aminoacyl-tRNA synthetase/tRNA pairs are crucial for the incorporation of unnatural amino acids in a site-specific manner. Here the authors use rational chimera design to create multiple efficient pairs that function in bacterial and mammalian systems for genetic code expansion.