Optimization of the 5ʹ untranslated region of mRNA vaccines

• Published in: Scientific reports Vol. 14; no. 1; pp. 19845 - 9
• Main Authors: Ma, Qi, Zhang, Xiaoguang, Yang, Jing, Li, Hongxia, Hao, Yanzhe, Feng, Xia
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 27.08.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 5' Untranslated Regions; 631/250/590; 631/337; Animals; Antigens; beta-Globins - genetics; COVID-19 vaccines; Cytochrome; Disease control; Efficiency; Fluorescence; Gene expression; Genes, Reporter; HEK293 Cells; HIV vaccine; Humanities and Social Sciences; Humans; Immunogenicity; In vitro transcription; mRNA Vaccines; multidisciplinary; Pathogens; Plasmids; Reporter gene; Reporter gene expression system; RNA, Messenger - genetics; Science; Science (multidisciplinary); Untranslated regions; Vaccines; Untranslated regions; In vitro transcription; mRNA vaccines; HIV vaccine; Reporter gene expression system
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-70792-x

To investigate the impact of different 5ʹ untranslated regions (UTRs) on mRNA vaccine translation efficiency, five dual-reporter gene expression plasmids with different 5ʹUTRs were constructed. The corresponding mRNA transcripts were transcribed and capped in vitro. By comparing the expression levels of reporter genes with different 5ʹUTRs, we identified the 5ʹUTR associated with the highest expression level. Subsequently, HIVgp145 mRNA vaccines containing various 5ʹUTRs were constructed and verified. The results demonstrated that mRNA 3 (β-globin 5ʹUTR) displayed the greatest number of green fluorescence-positive cells and the highest luciferase fluorescence intensity in the reporter gene expression system. Further, among the HIVgp145 mRNA vaccines with different 5ʹUTRs, mRNA 7 (β-globin 5ʹUTR) exhibited the highest level of expression. These findings indicate that it is feasible to use the 5ʹUTR of β-globin in an mRNA vaccine, laying the foundation for animal immunogenicity testing.