...homologous the off-target sites are compared to the on-target site, the specific site sequence, and the concentration...pX335 with BbsI/AgeI double digestion. As a successful insertion will destroy the BbsI sites, a double ...putting in the full target site in the donor. For example, choosing target sites that span the knock-in gene...which will together introduce cleavage of the target site with efficiency similar to using a single chimeric...promoter prefers a 'G' at the transcription start site to have high expression, so adding this G could ...breaks (DSBs) induced, compared to where the target site sequence (protospacer+PAM) is located? If there ...immediately adjacent to the nearly-homologous target sites. The mere presence of additional PAM sequences should...