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 this system optimized for mammals is currently used for synthetic gene therapy and C3P3 bioproduction tools.
Can C3P3 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 and in vivo for mammalian. However, C3P3 is not an appropriate technique for in vitro production of synthetic mRNA in reaction buffers.
Can C3P3 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 the purpose of genetic inhibition (shRNA, miRNA, lncRNA, ribozymes, aptamers) and possibly also appropriate for genome engineering by the editing system.
Are the modification of mRNA synthesized by the C3P3 system native?
Yes, the C3P3 system generates mRNA with native modifications. mRNA of eukaryotes undergoes multiple posttranscriptional modifications required for its biological activity. The C3P3 system generates mRNA modifications which are the same as those performed by the mammalian cells .
What are the performances of the C3P3 system in cellulo?
The performances of the C3P3-G3 system depends on the cell type and whether DNA contains or not CpG dinucleotides (CpG-free DNA is used for therapeutic applications). For instance, in human HEK-293 cells, C3P3 is 4-5 fold and 50-100-fold more efficient than standard expression system using CpG-rich or CpG-free DNA templates, respectively.
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