Can I use the Edit-R synthetic RNA components with my own Cas9 nuclease mRNA? Or mutant Cas9 nuclease plasmids?
We have validated the use of Edit-R synthetic tracrRNA and crRNAs and achieved efficient gene editing utilizing the Edit-R Cas9 expression plasmids in mammalian cell lines. Thus we cannot predict the efficacy of, nor can we troubleshoot experiments performed with, a mutant Cas9 nuclease expression vector or Cas9 mRNA. However, the repeat component of the crRNA sequence and the entire tracrRNA sequence are derived from the Streptococcus pyogenes CRISPR-Cas9 system, so they very likely can be used with another S. pyogenes-derived Cas9 component that is suitably optimized and sufficiently generates active Cas9 protein. Additionally, you must be able to efficiently co-transfect your Cas9 mRNA or plasmid DNA with the synthetic RNAs.