National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Street #100, Xicheng District, Beijing, 100052, China.
Sci Rep. 2024 Aug 27;14(1):19845. doi: 10.1038/s41598-024-70792-x.
To investigate the impact of different 5' untranslated regions (UTRs) on mRNA vaccine translation efficiency, five dual-reporter gene expression plasmids with different 5'UTRs were constructed. The corresponding mRNA transcripts were transcribed and capped in vitro. By comparing the expression levels of reporter genes with different 5'UTRs, we identified the 5'UTR associated with the highest expression level. Subsequently, HIVgp145 mRNA vaccines containing various 5'UTRs were constructed and verified. The results demonstrated that mRNA 3 (β-globin 5'UTR) 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 5'UTRs, mRNA 7 (β-globin 5'UTR) exhibited the highest level of expression. These findings indicate that it is feasible to use the 5'UTR of β-globin in an mRNA vaccine, laying the foundation for animal immunogenicity testing.
为了研究不同 5'非翻译区（UTR）对 mRNA 疫苗翻译效率的影响，构建了五个具有不同 5'UTR 的双报告基因表达质粒。体外转录并加帽相应的 mRNA 转录本。通过比较具有不同 5'UTR 的报告基因的表达水平，我们确定了与最高表达水平相关的 5'UTR。随后，构建并验证了含有各种 5'UTR 的 HIVgp145 mRNA 疫苗。结果表明，在报告基因表达系统中，mRNA3（β-珠蛋白 5'UTR）显示出最多的绿色荧光阳性细胞和最高的荧光素酶荧光强度。此外，在具有不同 5'UTR 的 HIVgp145 mRNA 疫苗中，mRNA7（β-珠蛋白 5'UTR）表达水平最高。这些发现表明，在 mRNA 疫苗中使用β-珠蛋白的 5'UTR 是可行的，为动物免疫原性测试奠定了基础。
