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. 2019 Feb 6;47(5):2681–2698. doi: 10.1093/nar/gkz069

Figure 1.

Figure 1.

The Firefly Luciferase gene reporter assay used for the optimization of the C3P3-G1 system. A diagram depicts the steps of gene expression from the C3P3-G1 expression system, using Firefly Luciferase as expression gene reporter. C3P3-G1 enzyme and T7 promoter-Luciferase plasmids (pT7φ10-Luciferase) are co-transfected into HEK-293 or CHO-K1 cultured cells. C3P3-G1 mRNA are expressed by the RNA polymerase II-dependent IE1 human CMV promoter/enhancer. The transcripts are subsequently translated into C3P3-G1 protein, which accumulates in the cell cytoplasm where it mediates transcription and capping of luciferase mRNAs from the pT7φ10-Luciferase plasmid, while polyadenylation is encoded in a poly[A]-track from the pT7φ10-Luciferase plasmid. Ultimately, luciferase protein is produced by mRNA translation, which is monitored by luciferin oxidation assay.