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Review
. 2024 May 22;124(10):6444-6500.
doi: 10.1021/acs.chemrev.3c00894. Epub 2024 Apr 30.

Engineering tRNAs for the Ribosomal Translation of Non-proteinogenic Monomers

Maxwell Sigal  1 , Satomi Matsumoto  1 , Adam Beattie  1 , Takayuki Katoh  1 , Hiroaki Suga  1
Affiliations
Review

Engineering tRNAs for the Ribosomal Translation of Non-proteinogenic Monomers

Maxwell Sigal et al. Chem Rev. .

Abstract

Ribosome-dependent protein biosynthesis is an essential cellular process mediated by transfer RNAs (tRNAs). Generally, ribosomally synthesized proteins are limited to the 22 proteinogenic amino acids (pAAs: 20 l-α-amino acids present in the standard genetic code, selenocysteine, and pyrrolysine). However, engineering tRNAs for the ribosomal incorporation of non-proteinogenic monomers (npMs) as building blocks has led to the creation of unique polypeptides with broad applications in cellular biology, material science, spectroscopy, and pharmaceuticals. Ribosomal polymerization of these engineered polypeptides presents a variety of challenges for biochemists, as translation efficiency and fidelity is often insufficient when employing npMs. In this Review, we will focus on the methodologies for engineering tRNAs to overcome these issues and explore recent advances both in vitro and in vivo. These efforts include increasing orthogonality, recruiting essential translation factors, and creation of expanded genetic codes. After our review on the biochemical optimizations of tRNAs, we provide examples of their use in genetic code manipulation, with a focus on the in vitro discovery of bioactive macrocyclic peptides containing npMs. Finally, an analysis of the current state of tRNA engineering is presented, along with existing challenges and future perspectives for the field.

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Conflict of interest statement

The authors declare no competing financial interest.

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