A Highly Optimized Human In vitro Translation System

Overview

Journal Cell Rep Methods

Specialty Cell Biology

Date 2024 Apr 12

PMID 38608690

Authors

Adrian Bothe

Nenad Ban

Affiliations

Soon will be listed here.

Abstract

In vitro translation is an important method for studying fundamental aspects of co- and post-translational gene regulation, as well as for protein expression in the laboratory and on an industrial scale. Here, by re-examining and improving a human in vitro translation system (HITS), we were able to develop a minimal system where only four components are needed to supplement human cell lysates. Functional characterization of our improved HITS revealed the synergistic effect of mRNA capping and polyadenylation. Furthermore, we found that mRNAs are translated with an efficiency equal to or higher than existing state-of-the-art mammalian in vitro translation systems. Lastly, we present an easy preparation procedure for cytoplasmic extracts from cultured HeLa cells, which can be performed in any cell culture laboratory. These methodological advances will allow HITSs to become a widespread tool in basic molecular biology research.

Citing Articles

Optimizing Human Cell-Free System for Efficient Protein Production.

Mansour A, Kipper K, Pulk A J Microbiol Biotechnol. 2025; 35:e2410026.

PMID: 40016147 PMC: 11896798. DOI: 10.4014/jmb.2410.10026.

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