Schematic of the split nLuc Tornado translation system and the circular split nLuc reporter assay. (A) Construct design of the Tornado translation system using a split nLuc ORF. The circular split nLuc reporter comprises an SmBiT (pink), followed by an IRES (green), followed by an LgBiT (blue). When the RNA is transcribed, ribozyme (orange) cleavage and ligation will occur. The split nLuc protein will be translated by IRES translation. (B) Luminescence from circular split nLuc system is dependent on circularization. Luminescence from HEK293T cells transfected with plasmids encoding the circular split nLuc reporter with either a mutant ribozyme (mutTornado CVB3 IRES) or a functional ribozyme (CVB3 IRES). (CJ) Circularization junction.
