Some answers to most frequent questions on the C3P3 artificial expression system and its applications
How was developped the C3P3 expression system?
The C3P3 system is molecular biorector able to synthesize autonomously fully mature mRNA. This system contains a several modules, which ensure the production of the mRNA chain and its multiple modifications required for its biological activity in a coordinated way. C3P3 has been entirely developed, module per module, by synthetic biology and molecular engineering.
What generation of the C3P3 expression system is currently used?
The improvement of the C3P3 system is a continuous process. Each next generation improves its performances. The third generation of this system is currently used for all synthetic gene therapy applications, as well as tools for bioproduction of recombinant proteins and viruses.
Is C3P3 is the first ever non-viral synthetic transcription system for eukaryotes?
Yes, to to the best of our knowledge, the C3P3 expression system is the first ever eukaryotic non-viral artificial expression system. C3P3 has been developed from scratch by Eukarÿs through a complex process including multiple biological engineering steps. The third generation (C3P3-G3) of the C3P3 system optimized for mammals is currently used for synthetic gene therapy and C3P3 bioproduction tools.
Can the C3P3 system be used of in cellulo or in vivo applications? What about in vitro synthesis of mRNA with C3P3?
Yes, the C3P3 system synthesizes the mRNA(s) of interest in the cytoplasm of “living cells”. C3P3 has demonstrated its efficacy both in cellulo in cultured cells and in vivo for mammalian. However, C3P3 is not an appropriate technique for in vitro production of synthetic mRNA in reaction buffers.
Can C3P3 system can be also used for gene expression repression?
Yes, the C3P3 system also allows the production of non-coding RNAs which can be used for genetic expression inhibition (shRNA, miRNA, lncRNA, ribozymes, aptamers) and possibly also for genome engineering by the editing system.
Are the modifications of mRNA synthesized by the C3P3 system native?
Eukaryotic mRNA undergoes multiple post-transcriptional modifications, which are required for biological activity. The C3P3 system generates mRNA molecules with native modifications which are the structurally the same as those performed by the mammalian cells.
What are the performances of the C3P3 system in cellulo in cultured cells?
The current performances of the C3P3-G3 system depends on the cell type and whether DNA templates contains or not CpG dinucleotides (CpG-free DNA is used for therapeutic applications). For instance, in human HEK-293 cells, the C3P3 system is 4-5 fold and 20-30-fold more efficient than standard expression system using DNA templates with or without CpG dinucleotides, respectively.