Investigation of the effect of 5’UTR on immunogenicity in coxsackievirus VP1 mRNA vaccine

Abstract Hand, foot and mouth disease (HFMD) is a group of common infectious diseases caused by various enteroviruses, which are transmitted through gastrointestinal tract, respiratory tract and contact. The mRNA vaccine is safe, effective, simple to produce and easy to promote, and has attracted much attention in recent years. Starting from the screening modification of the 5’UTR of antigen nucleic acid sequence, the mRNA translation efficiency was improved by introducing the antigen epitope cleavage processing signal through the untranslated region screening modification, and the nucleic acid vaccine design with stable translation characteristics and effective control of inflammation homeostasis was studied to provide a reference for the molecular design of future mRNA vaccines. In this study, 5’UTRs of different lengths of human β-globin origin were successfully constructed in front of the gene sequences of CA6-VP1 and CA10-VP1; the target gene sequences were transcribed in vitro with capping and tailing reactions to obtain structurally complete mRNA sequences; 293T cells were transfected and Western Blot assays were performed to find that the target genes with the modified 5’UTRs attached The mRNA was able to express the protein correctly and stably, and the protein expression was more compared with the original sequence without the modified 5’UTR. The human β-globin 5’UTR is useful for improving the translation efficiency and protein stability of the target gene mRNA. Further experimental evidence is needed to assess its induction of immune responses in vivo.