Table 2.
Gene editing | Number of analyzed off-targets | Number of analyzed clones | Number of SVs and long indels (×10−3/off-target/clone) at regions surrounding the predicted off-target sites | ||||
---|---|---|---|---|---|---|---|
±10 bp | ±25 bp | ±50 bp | ±100 bp | ||||
sgEx4_mt20s + sgS14 [Nickase] |
No treatment | 198 | 12 | 0.00 | 0.00 | 0.00 | 0.00 |
Gene corrected | 198 | 13 | 0.00 | 0.00 | 0.00 | 0.00 | |
sgEx4_mt20s + sgS8 [Nickase] |
No treatment | 198 | 12 | 0.00 | 0.00 | 0.00 | 0.00 |
Gene corrected | 198 | 10 | 0.00 | 0.00 | 0.00 | 0.00 | |
sgEx4_mt20s + sgS34 [Nickase] |
No treatment | 198 | 12 | 0.00 | 0.00 | 0.00 | 0.00 |
Gene corrected | 198 | 9 | 0.00 | 0.56 | 0.56 | 0.56 | |
sgEx4_mt20s [Cas9] + ssODN |
No treatment | 99 | 12 | 0.00 | 0.00 | 0.00 | 0.00 |
Gene corrected | 99 | 15 | 2.00 | 3.33 | 3.33 | 4.00 |
The number of SVs and indels (×10−3/off-target/clone) in the ±10, ±25, ±50, and ±100-bp regions surrounding the 3′ end of the off-target sequences was analyzed using the MANTA method. Regions surrounding the 99 off-target sites of each sgRNA predicted in silico were analyzed. SV, structural variant; WGS, whole Genome Sequencing; SCC, single-cell derived clone.