I am working against a series of genetic barriers to transformation in a bacteria which has never been successfully transformed
The genome shows the presence of an endogenous Type-II Crispr system which has an array of 14 spacers.
If I align these spacers with my plasmid of interest there is some pretty high levels of homology, not exact, but sometimes 100% identity across 60% of the spacer
Does anyone know how specific does the spacer/foreign sequence match have to be to be degraded by the endonuclease domain?
For example, if there is a few interspersed single bp differences, will this still be degraded? (like a primer/template relationship in PCR)
Also has anyone successfully used a crispr inhibiting protein (gene from phage) cloned to a plasmid to overcome a Crispr system?
Thanks in advance