Optimization of the 5ʹ untranslated region of mRNA vaccines

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 level...

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Bibliographic Details
Published in:Scientific reports 2024-08, Vol.14 (1), p.19845-9
Main Authors: Ma, Qi, Zhang, Xiaoguang, Yang, Jing, Li, Hongxia, Hao, Yanzhe, Feng, Xia
Format: Article
Language:English
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
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-024-70792-x