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Chimeric design of pyrrolysyl-tRNA synthetase/tRNA pairs and canonical synthetase/tRNA pairs for genetic code expansion

Author

Listed:
  • Wenlong Ding

    (Zhejiang University)

  • Hongxia Zhao

    (Zhejiang University)

  • Yulin Chen

    (Zhejiang University)

  • Bin Zhang

    (Zhejiang University)

  • Yang Yang

    (Nanjing University)

  • Jia Zang

    (Zhejiang University)

  • Jing Wu

    (Zhejiang University)

  • Shixian Lin

    (Zhejiang University)

Abstract

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.

Suggested Citation

  • Wenlong Ding & Hongxia Zhao & Yulin Chen & Bin Zhang & Yang Yang & Jia Zang & Jing Wu & Shixian Lin, 2020. "Chimeric design of pyrrolysyl-tRNA synthetase/tRNA pairs and canonical synthetase/tRNA pairs for genetic code expansion," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-16898-y
    DOI: 10.1038/s41467-020-16898-y
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    Cited by:

    1. Hongxia Zhao & Wenlong Ding & Jia Zang & Yang Yang & Chao Liu & Linzhen Hu & Yulin Chen & Guanglong Liu & Yu Fang & Ying Yuan & Shixian Lin, 2021. "Directed-evolution of translation system for efficient unnatural amino acids incorporation and generalizable synthetic auxotroph construction," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
    2. Dan-Dan Liu & Wenlong Ding & Jin-Tao Cheng & Qiushi Wei & Yinuo Lin & Tian-Yi Zhu & Jing Tian & Ke Sun & Long Zhang & Peilong Lu & Fan Yang & Chao Liu & Shibing Tang & Bing Yang, 2024. "Characterize direct protein interactions with enrichable, cleavable and latent bioreactive unnatural amino acids," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

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