Fig. 6

The strategy to exclude the off-target effects in the descendants of the engineering mice based on the CRISPR/Cas9 system. To avoid the off-target effects in the CRISPR mice, the following strategy is performed. Step 1) BLAST search the potential matching loci in mouse genome with the gRNA applied for mouse engineering; Step 2) Amplify and sequence the potential off-target loci with specific primers in generation 1 mice; Step 3) Select those mice that do have off-target effects to be further studied. The strategy can be applied on any mouse constructed via the CRISPR/Cas9 system including Non-homologous end joining, NHEJ-drive knockout (frame shift mutation) and Homology directed repair, HDR-drive knock-in (donor replacement) mice