How to mutate sgRNAs to prevent re-cutting

We often want to mutate sgRNA target sites in HDR donor templates or complemented constructs so that they are not targeted by the sgRNA that cut the wild type region.

The best way to do this is to either mutate the PAM site (GG), or if this not possible, to mutate the 1-5bp seed region proximal to the PAM site (see discussion in Zhang 2015).