TY  - JOUR
IS  - 1
N2  - The manufacturing of mRNA vaccines relies on cell-free based systems that are easily scalable and flexible compared with the traditional vaccine manufacturing processes. Typically, standard processes yield 2 to 5 g L?1 of mRNA, with recent process optimisations increasing yields to 12 g L?1. However, increasing yields can lead to an increase in the production of unwanted by-products, namely dsRNA. It is therefore imperative to reduce dsRNA to residual levels in order to avoid intensive purification steps, enabling cost-effective manufacturing processes. In this work, we exploit sequence modifications downstream of the T7 RNA polymerase promoter to increase mRNA yields whilst simultaneously minimising dsRNA. In particular, transcription performance was optimised by modifying the sequence downstream of the T7 promoter with additional AT-rich sequences. We have identified variants that were able to produce higher amounts of mRNA (up to 14 g L?1) in 45 min of reaction. These variants exhibited up to a 30% reduction in dsRNA byproduct levels compared to a wildtype T7 promoter, and have similar EGFP protein expression. The results show that optimising the non-coding regions can have an impact on mRNA production yields and quality, reducing overall manufacturing costs.
ID  - discovery10191433
UR  - http://dx.doi.org/10.1038/s41598-024-59978-5
TI  - Comprehensive evaluation of T7 promoter for enhanced yield and quality in mRNA production
Y1  - 2024///
AV  - public
KW  - Nucleic-acid therapeutics
KW  -  RNA
KW  -  Synthetic biology
A1  - Sari, Yustika
A1  - Sousa Rosa, Sara
A1  - Jeffries, Jack
A1  - Marques, Marco PC
PB  - Springer Science and Business Media LLC
JF  - Scientific Reports
N1  - © 2024 Springer Nature Limited. This article is licensed under a Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/).
VL  - 14
ER  -