Extensive breaking of genetic code degeneracy with non-canonical amino acids

Genetic code expansion (GCE) offers many exciting opportunities for the creation of synthetic organisms and for drug discovery methods that utilize in vitro translation. One type of GCE, sense codon reassignment (SCR), focuses on breaking the degeneracy of the 61 sense codons which encode for only 2...

Full description

Saved in:
Bibliographic Details
Published in:Nature communications 2023-08, Vol.14 (1), p.5008-5008, Article 5008
Main Authors: McFeely, Clinton A. L., Shakya, Bipasana, Makovsky, Chelsea A., Haney, Aidan K., Ashton Cropp, T., Hartman, Matthew C. T.
Format: Article
Language:English
Subjects:
631/337/574/1789
631/337/574/1793
631/553/552
631/92/611
639/638/92/610
82/58
Amino acids
Amino Acids - genetics
Biological Assay
Codons
Drug Discovery
Genetic Code
Humanities and Social Sciences
In vitro methods and tests
Language
Magnoliopsida
multidisciplinary
Post-transcription
Ribosomes
RNA, Transfer, Amino Acyl
Science
Science (multidisciplinary)
Transfer RNA
tRNA
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Genetic code expansion (GCE) offers many exciting opportunities for the creation of synthetic organisms and for drug discovery methods that utilize in vitro translation. One type of GCE, sense codon reassignment (SCR), focuses on breaking the degeneracy of the 61 sense codons which encode for only 20 amino acids. SCR has great potential for genetic code expansion, but extensive SCR is limited by the post-transcriptional modifications on tRNAs and wobble reading of these tRNAs by the ribosome. To better understand codon-tRNA pairing, here we develop an assay to evaluate the ability of aminoacyl-tRNAs to compete with each other for a given codon. We then show that hyperaccurate ribosome mutants demonstrate reduced wobble reading, and when paired with unmodified tRNAs lead to extensive and predictable SCR. Together, we encode seven distinct amino acids across nine codons spanning just two codon boxes, thereby demonstrating that the genetic code hosts far more re-assignable space than previously expected, opening the door to extensive genetic code engineering. Genetic code expansion is limited by the degeneracy of the 61 sense codons which encode for only 20 amino acids. Here, the authors show that by combining hyperaccurate ribosomes and in vitro transcribed tRNAs, dramatic and extensive breaking of sense codon degeneracy can be achieved.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-023-40529-x