3’ to 5’ translation of linear mRNAs?

3’→5’ translation (termed “backward translation”) mediated by eukaryotic circular RNAs has been reported recently. Whether linear mRNA can produce proteins through backward translation remained elusive. Here, we demonstrate that linear mRNAs can mediate backward translation in vitro . Backward translation was detected in human cells transfected with expression vectors used to produce linear mRNAs. Importantly, synthetic linear mRNAs could produce backward translation signal in multiple in vitro translation systems, with a level comparable to that of conventional 5’ to 3’ forward translation. Furthermore, we demonstrated that a single translation initiating sequence (TIS) was able to drive backward and forward translation in an in vitro experimental setup, with the efficiency of backward translation found to be significant lower than that of forward translation. In summary, this study further reinforces the notion of translation flexibility, demonstrates a broader applicability of backward translation and heralds a new strategy in synthetic biology.