TRANSLATION ENHANCEMENT OF IN-VITRO-TRANSCRIBED MESSENGER RNA BY INFLUENZA A VIRUS DERIVED NON-STRUCTURAL PROTEIN 1

Abstract

In this thesis, we show our exploration of applying influenza A virus derived non-structural protein 1 (NS1) to modulate immunogenicity and translation capacity of in vitro-transcribed (IVT) mRNA. Co-transfection of mRNA encoding NS1 and reporter gene mRNA enhanced reporter gene expression by over 10 times. NS1’s inhibition of interferon regulatory factor 3 (IRF3), protein kinase RNA-activated (PKR), and most importantly cleavage and polyadenylation factor 30kDa (CPSF30) was found to be contributing mechanism. One single intramuscular co-injection of NS1 mRNA and reporter gene mRNA was shown to mediate great enhancement of reporter gene expression and prolong the kinetics of expression by up to 7 days. In a protein therapy model, co-injection of NS1 mRNA and erythropoietin mRNA significantly increased hematocrit of treated mice. In a vaccination model, co-injection of NS1 mRNA, interferon mRNA and antigen mRNA not only promised sustained antigen expression, but also induced stronger antibody response against antigen.