A Streamlined In Vitro mRNA Production Evaluation for mRNA-Based Vaccines and Therapeutics.

To develop an effective strategy for in vitro mRNA production, it is crucial to evaluate the efficiency of the in vitro transcription platform. This can be accomplished using reporter genes, such as the luciferase-encoding gene. Luciferase activity assays provide a reliable means to assess the translation efficiency of in vitro transcribed mRNAs and to explore molecular dynamics associated with untranslated regions, capping, nucleotide analog incorporation, polyadenylation, and codon usage optimization. In this study, we propose a novel approach to performing the luciferase assay, offering a simpler, faster, and high-throughput method for evaluating in vitro generated transcripts to be employed for veterinary and human vaccine purposes as well as mRNA therapeutics.