Overview of ovoD-assisted gene targeting efficiency and precision of the arm locus without and with the use of modified PAM sites
| Allele . | With unmodified PAM site in HDR plasmid, n/N (%) . | With modified PAM site in HDR plasmid, n/N (%) . |
|---|---|---|
| armWT | ||
| No. of clonal F1 offspring analyzed | 8 | 14 |
| With edited missense mutation | 0/8 (0) | 3/14 (21) |
| With correct unmodified/modified 5′-PAM site sequence | − | 3/3 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 3/3 (100) |
| arm#1 | ||
| No. of clonal F1 offspring analyzed | 13 | NA |
| With edited missense mutation | 0/13 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#2 | ||
| No. of clonal F1 offspring analyzed | 2 | 18 |
| With edited missense mutation | 0/2 (0) | 12/18 (67) |
| With correct unmodified/modified 5′-PAM site sequence | − | 12/12 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 12/12 (100) |
| arm#3 | ||
| No. of clonal F1 offspring analyzed | 8 | NA |
| With edited missense mutation | 0/8 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#4 | ||
| No. of clonal F1 offspring analyzed | 7 | NA |
| With edited missense mutation | 0/7 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − |
| Allele . | With unmodified PAM site in HDR plasmid, n/N (%) . | With modified PAM site in HDR plasmid, n/N (%) . |
|---|---|---|
| armWT | ||
| No. of clonal F1 offspring analyzed | 8 | 14 |
| With edited missense mutation | 0/8 (0) | 3/14 (21) |
| With correct unmodified/modified 5′-PAM site sequence | − | 3/3 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 3/3 (100) |
| arm#1 | ||
| No. of clonal F1 offspring analyzed | 13 | NA |
| With edited missense mutation | 0/13 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#2 | ||
| No. of clonal F1 offspring analyzed | 2 | 18 |
| With edited missense mutation | 0/2 (0) | 12/18 (67) |
| With correct unmodified/modified 5′-PAM site sequence | − | 12/12 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 12/12 (100) |
| arm#3 | ||
| No. of clonal F1 offspring analyzed | 8 | NA |
| With edited missense mutation | 0/8 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#4 | ||
| No. of clonal F1 offspring analyzed | 7 | NA |
| With edited missense mutation | 0/7 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − |
Notes: For experimental sets using unmodified PAM sites, “correct” refers to their wild-type sequence, and for experiments using modified PAM sites, “correct” refers to the modified sequence.
Overview of ovoD-assisted gene targeting efficiency and precision of the arm locus without and with the use of modified PAM sites
| Allele . | With unmodified PAM site in HDR plasmid, n/N (%) . | With modified PAM site in HDR plasmid, n/N (%) . |
|---|---|---|
| armWT | ||
| No. of clonal F1 offspring analyzed | 8 | 14 |
| With edited missense mutation | 0/8 (0) | 3/14 (21) |
| With correct unmodified/modified 5′-PAM site sequence | − | 3/3 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 3/3 (100) |
| arm#1 | ||
| No. of clonal F1 offspring analyzed | 13 | NA |
| With edited missense mutation | 0/13 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#2 | ||
| No. of clonal F1 offspring analyzed | 2 | 18 |
| With edited missense mutation | 0/2 (0) | 12/18 (67) |
| With correct unmodified/modified 5′-PAM site sequence | − | 12/12 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 12/12 (100) |
| arm#3 | ||
| No. of clonal F1 offspring analyzed | 8 | NA |
| With edited missense mutation | 0/8 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#4 | ||
| No. of clonal F1 offspring analyzed | 7 | NA |
| With edited missense mutation | 0/7 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − |
| Allele . | With unmodified PAM site in HDR plasmid, n/N (%) . | With modified PAM site in HDR plasmid, n/N (%) . |
|---|---|---|
| armWT | ||
| No. of clonal F1 offspring analyzed | 8 | 14 |
| With edited missense mutation | 0/8 (0) | 3/14 (21) |
| With correct unmodified/modified 5′-PAM site sequence | − | 3/3 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 3/3 (100) |
| arm#1 | ||
| No. of clonal F1 offspring analyzed | 13 | NA |
| With edited missense mutation | 0/13 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#2 | ||
| No. of clonal F1 offspring analyzed | 2 | 18 |
| With edited missense mutation | 0/2 (0) | 12/18 (67) |
| With correct unmodified/modified 5′-PAM site sequence | − | 12/12 (100) |
| With correct unmodified/modified 3′-PAM site sequence | − | 12/12 (100) |
| arm#3 | ||
| No. of clonal F1 offspring analyzed | 8 | NA |
| With edited missense mutation | 0/8 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − | |
| arm#4 | ||
| No. of clonal F1 offspring analyzed | 7 | NA |
| With edited missense mutation | 0/7 (0) | |
| With correct unmodified/modified 5′-PAM site sequence | − | |
| With correct unmodified/modified 3′-PAM site sequence | − |
Notes: For experimental sets using unmodified PAM sites, “correct” refers to their wild-type sequence, and for experiments using modified PAM sites, “correct” refers to the modified sequence.
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