I am designing a CRISPR experiment to correct a point mutation in iPSCs.
I will use a plasmid encoding gRNA, Cas9, GFP/Puro and a ssODN as repair template.
Regarding the gRNA I don´t have lot of options: one cuts very close to the insertion site (2 bp) but has medium on/off target score while the others have higher on/off target score but further away from the insertion site (15-20 bp).
In your personal experience what would you prioritize in such a case? gRNA score or distance from insertion site?
Any comment and suggestion is welcomed.