Basavaraj Sajjanar, Shreya Gopinath, Prasanth Nadarajan, Vishnu Kiran Marreddy, Narayanan Krishnaswamy, Viswas K Nagaleekar, Madhusudan Hosamani, B P Sreenivasa, Pallab Chaudhuri, Suresh H Basagoudanavar
Synthetic messenger RNA (mRNA) is emerging as next generation vaccine platform. As natural 5'UTRs of particular genes do not work for other than its own mRNA, it is essential to identify an ideal 5'UTR supporting higher translation efficiency for different antigens. Six natural 5'UTRs derived from α, β-globin genes of host animals and other viral genes were cloned upstream to the coding sequences of reporter proteins. The corresponding capped mRNA were produced through in vitro transcription and the translation efficiency was assessed using flow cytometry, bioluminescence, immunofluorescence and western blot experiments. The results revealed higher translation of reporter proteins by bovine β-globin 5'UTR in different cell lines. Further, the chosen 5'UTR supported mRNA translation to produce different antigenic proteins such as rabies virus glycoprotein (RVG), major structural protein of foot and mouth disease virus (VP1) and bacterial epsilon toxoid protein (Eptx). Bovine -globin 5'UTR not only increased the translation efficiency of reporter proteins but also supported the expression of different antigenic proteins. Hence, bovine -globin 5'UTR can be used in the mRNA vaccine designs against animal diseases.