C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system

Abstract : Most eukaryotic expression systems make use of host-cell nuclear transcriptional and post-transcriptional machineries. Here, we present the first generation of the chimeric cytoplasmic capping-prone phage polymerase (C3P3-G1) expression system developed by biological engineering, which generates capped and polyadenylated transcripts in host-cell cytoplasm by means of two components. First, an artificial single-unit chimeric enzyme made by fusing an mRNA capping enzyme and a DNA-dependent RNA polymerase. Second, specific DNA templates designed to operate with the C3P3-G1 enzyme , which encode for the transcripts and their artificial polyadenylation. This system, which can potentially be adapted to any in cellulo or in vivo eu-karyotic expression applications, was optimized for transient expression in mammalian cells. C3P3-G1 shows promising results for protein production in Chinese Hamster Ovary (CHO-K1) cells. This work also provides avenues for enhancing the performances for next generation C3P3 systems.
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Philippe Jaïs, Etienne Decroly, Eric Jacquet, Marine Le boulch, Aurélien Jaïs, et al.. C3P3-G1: first generation of a eukaryotic artificial cytoplasmic expression system. Nucleic Acids Research, Oxford University Press, 2019, 47 (5), pp.2681-2698. ⟨10.1093/nar/gkz069⟩. ⟨inserm-02016614⟩

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