Reprogramming the Genetic Code: From Triplet to Quadruplet Codes

The genetic code of cells is near‐universally triplet, and since many ribosomal mutations are lethal, changing the cellular ribosome to read nontriplet codes is challenging. Herein we review work on the incorporation of unnatural amino acids into proteins in response to quadruplet codons, and the creation of an orthogonal translation system in the cell that uses an evolved orthogonal ribosome to efficiently direct the incorporation of unnatural amino acids in response to quadruplet codons. Using this system multiple distinct unnatural amino acids have been incorporated and used to genetically program emergent properties into recombinant proteins. Extension of approaches to incorporate multiple unnatural amino acids may allow the combinatorial biosynthesis of materials and therapeutics, and drive investigations into whether life with additional genetically encoded polymers can evolve to perform functions that natural biological systems cannot. On all fours: Recent work has used quadruplet codons to encode unnatural amino acids. Multiple unnatural amino acids have been incorporated into proteins through orthogonal protein translation using a quadruplet decoding ribosome (see figure; mutations to ribosome are shown in red).