Selected article for: "luciferase expression and translational efficiency"
Author: Jaïs, Philippe H; Decroly, Etienne; Jacquet, Eric; Le Boulch, Marine; Jaïs, Aurélien; Jean-Jean, Olivier; Eaton, Heather; Ponien, Prishila; Verdier, Fréderique; Canard, Bruno; Goncalves, Sergio; Chiron, Stéphane; Le Gall, Maude; Mayeux, Patrick; Shmulevitz, Maya
Title: C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system
  • Document date: 2019_3_18
    • ID: 6nq7y1qe_52
    Snippet: We sought to recapitulate the main components of the vaccinia virus-T7RNAP expression system in a non-viral plasmid-based expression system, therefore devoid of any risk of transmitting infection and virus-mediated cytotoxicity. The vaccinia virus capping enzyme consists of a 97and a 33-kDa subunit that are encoded by the vaccinia virus-D1R and D12L genes respectively (32) . The D1R product has enzymatic activities (33) (34) (35) , whereas the D1.....
    Document: We sought to recapitulate the main components of the vaccinia virus-T7RNAP expression system in a non-viral plasmid-based expression system, therefore devoid of any risk of transmitting infection and virus-mediated cytotoxicity. The vaccinia virus capping enzyme consists of a 97and a 33-kDa subunit that are encoded by the vaccinia virus-D1R and D12L genes respectively (32) . The D1R product has enzymatic activities (33) (34) (35) , whereas the D12L product acts as its allosteric regulator (36) . Both D1R and D12L were cloned into independent plasmids and cotransfected into HEK-293 cells in combination with pCMV-T7RNAP and pT710-Luciferase plasmids. Luciferase expression was increased by 9-fold in cells co-transfected with D1R and D12L plasmids in comparison to T7RNAP plasmid alone, which confirmed the importance of capping for T7-transcripts translational efficiency ( Figure 2 ).

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