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. 2022 Oct 18;19(1):1103–1114. doi: 10.1080/15476286.2022.2135299

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© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Evolution of orthogonal special ribosomes. (A) Early orthogonal specialised ribosomes were generated by random mutagenesis in the RNA-binding sequences (RBS) on mRNA and its complementary message-binding sequence (MBS) on rRNA in E. coli [134]. Orthologue ribosomes have been further evolved to increase the in vivo efficiency of unnatural amino acid incorporation (ribo-X [139]) and enable efficient incorporation of amber and quadruplet codons (ribo-Q1 [141,142].(B) The engineering of the fully orthogonal ribosomes became possible with the advent of the ribosome covalently linked through an optimised RNA staple (O-stapled ribosomes [147,148]) or tethered ribosomal subunits with RNA linker (Ribo-T/Ribo-T v2 [146–149];.