Table 2.

Summary of predicted off-target sites for sgRNA_U1 and sgRNA_D1, and off-target variations identified

Predicted off-target sites	With two mismatches	With three mismatches	With four mismatches	Number of off-target SNVs and indels
Exonic	0	0	8	0
Intergenic	0	6	122	0
Intronic	2	6	82	0

Summary of the predicted off-target sites with four or fewer mismatches for both sgRNA_U1 (total of 55 sites) and sgRNA_D1 (total of 173 sites) separated into genic type. Whole genome sequence analysis of the founder F₀ mouse, used to establish the line that was phenotyped in this study, demonstrated no modifications to be present at these 228 predicted off-target sites