Table 1.
Numbers of SNVs and indels evaluated by a GATK analysis of WGS of SCCs established before or after gene correction
| Gene editing | Number of analyzed off-targets | Number of analyzed clones | Number of SNVs and indels (×10-2/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 | 0.04 | 0.04 | 0.04 | 0.17 |
| Gene corrected | 198 | 13 | 0.00 | 0.00 | 0.00 | 0.04 | |
|
sgEx4_mt20s + sgS8 [Nickase] |
No treatment | 198 | 12 | 0.00 | 0.00 | 0.00 | 0.04 |
| Gene corrected | 198 | 10 | 0.00 | 0.00 | 0.00 | 0.20 | |
|
sgEx4_mt20s + sgS34 [Nickase] |
No treatment | 198 | 12 | 0.04 | 0.04 | 0.21 | 0.75 |
| Gene corrected | 198 | 9 | 0.00 | 0.00 | 0.00 | 0.22 | |
|
sgEx4_mt20s [Cas9] + ssODN |
No treatment | 99 | 12 | 0.00 | 0.00 | 0.00 | 0.00 |
| Gene corrected | 99 | 15 | 2.53 | 3.00 | 3.00 | 3.60 | |
The number of SNVs and indels (×10−2/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 GATK method. Regions surrounding the 99 off-target sites of each sgRNA predicted in silico were analyzed. SNV, single nucleotide variant; WGS, whole Genome Sequencing; SCC, single-cell derived clone.