Ser/Leu-swapped cell-free translation system constructed with natural/in vitro transcribed-hybrid tRNA set.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 38755134.
- Also identified by DOI 10.1038/s41467-024-48056-z and PMC identifier 11099018.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
The Ser/Leu-swapped genetic code can act as a genetic firewall, mitigating biohazard risks arising from horizontal gene transfer in genetically modified organisms. Our prior work demonstrated the orthogonality of this swapped code to the standard genetic code using a cell-free translation system comprised of 21 in vitro transcribed tRNAs. In this study, to advance this system for protein engineering, we introduce a natural/in vitro transcribed-hybrid tRNA set. This set combines natural tRNAs from Escherichia coli (excluding Ser, Leu, and Tyr) and in vitro transcribed tRNAs, encompassing anticodon-swapped tRNA<sup>Ser</sup><sub>GAG</sub> and tRNA<sup>Leu</sup><sub>GGA</sub>. This approach reduces the number of in vitro transcribed tRNAs required from 21 to only 4. In this optimized system, the production of a model protein, superfolder green fluorescent protein, increases to 3.5-fold. With this hybrid tRNA set, the Ser/Leu-swapped cell-free translation system will stand as a potent tool for protein production with reduced biohazard concerns in future biological endeavors.
Medical subject headings
- Cell-Free System
- Protein Biosynthesis
- Escherichia coli