Table 3. Multiple targets Cas9 and Cas12a candidates for ChCBF genes.
All possible genome targets and offtargets (with ARES-GT thresholds: L0 = 4 and L1 = 3) of each candidate are listed with indication of genome coordinates (Cardamine hirsuta v1.0) and whether it corresponds to a CBF gene. In alignments, black boxes mark mismatches and a space separates PAM (NGG/NAG or TTTN) from sequence. Differences in the “N” position in the PAM are not marked.
| Candidate ID | Targets + Offtargets (L0 = 4, L1 =3) | |||||
|---|---|---|---|---|---|---|
| C. hirsuta | Gene | chrom | start | end | sense | sequence |
| Cas9ChCBF1_004 | ChCBF2 | 4 | 6514798 | 6514820 | + | AGCTGTCCCAAGAAACCAGC TGG |
| ChCBF1 | 7 | 17908883 | 17908905 | - | CCG GCTGGTTTCTTGGGACAGCT | |
| Cas9ChCBF1_010 | ChCBF2 | 4 | 6514878 | 6514900 | + | CTCCGGTAAGTGGGTGTGTG AGG |
| ChCBF1 | 7 | 17908803 | 17908825 | - | CCT CACACACCCACTTACCGGAGE | |
| Cas9ChCBF1_018 | ChCBF2 | 4 | 6514910 | 6514932 | + | CAAACAAGAAATCTAGGATT TGG |
| ChCBF1 | 7 | 17908771 | 17908793 | - | CCA AATCCTAGATTTCTTGTTTG | |
| ChCBF3 | 8 | 13812274 | 13812296 | - | CCA AATCCTCGATTTCTTGTTAG | |
| ---- | 5 | 18638271 | 18638293 | - | CTT AATCCTACATTTGTAGTTTG | |
| ---- | 5 | 21152837 | 21152859 | - | CTT AATCCTACATTTCTGGTTTT | |
| Cas9ChCBF1_013 | ChCBF2 | 4 | 6514915 | 6514937 | + | AAGAAATCTAGGATTTGGCT TGG |
| ChCBF1 | 7 | 17908766 | 17908788 | - | CCG AGCCAAATCCTAGATTTCTT | |
| ---- | 8 | 18333140 | 18333162 | - | CCA AGCCAAATCCTAGAACCCTT | |
| ---- | 1 | 5556241 | 5556263 | + | AGGAAACGGAGGATTTGGCT TGG | |
| ---- | 1 | 370416 | 370438 | + | AAAAAATCTCGGATTTGGCT CGG | |
| ChCBF3 | 8 | 13812269 | 13812291 | - | CCT AACCAAATCCTCGATTTCTT | |
| Cas9ChCBF1_033 | ChCBF2 | 4 | 6515264 | 6515286 | + | TGCCGCCTCCGTCCGTACAA TGG |
| ChCBF1 | 7 | 17908390 | 17908412 | - | CCA TTGTACGGACGGAGGCGGCA | |
| NSCAFA. | 444 | 2316 | 2338 | + | CGCCGCCACCGTCCGTACAC CGG | |
| Cas9ChCBF1_036 | ChCBF2 | 4 | 6514793 | 6514815 | - | CCG TGAGCTGTCCCAAGAAACCA |
| ChCBF1 | 7 | 17908888 | 17908910 | + | TGGTTTCTTGGGACAGCTCA CGG | |
| Cas9ChCBF1_043 | ChCBF2 | 4 | 6514880 | 6514902 | - | CCG GTAAGTGGGTGTGTGAGGTA |
| ChCBF1 | 7 | 17908801 | 17908823 | + | TACCTCACACACCCACTTAC CGG | |
| Cas9ChCBF1_044 | ChCBF2 | 4 | 6514909 | 6514931 | - | CCA AACAAGAAATCTAGGATTTG |
| ChCBF1 | 7 | 17908772 | 17908794 | + | CAAATCCTAGATTTCTTGTT TGG | |
| ChCBF3 | 8 | 13812275 | 13812297 | + | CAAATCCTCGATTTCTTGTT AGG | |
| Cas9ChCBF1_056 | ChCBF2 | 4 | 6515266 | 6515288 | - | CCG CCTCCGTCCGTACAATGGAA |
| ChCBF1 | 7 | 17908388 | 17908410 | + | TTCCATTGTACGGACGGAGG CGG | |
| ---- | 2 | 8347578 | 8347600 | + | GGCCAGAGTACGGACGGAGG AGG | |
| Cas9ChCBF1_057 | ChCBF2 | 4 | 6515269 | 6515291 | - | CCT CCGTCCGTACAATGGAATCA |
| ChCBF1 | 7 | 17908385 | 17908407 | + | TGATTCCATTGTACGGACGG AGG | |
| ---- | 1 | 17089187 | 17089209 | + | TGGTCCGGTTGTACGGACGG CGG | |
| ---- | 5 | 5225681 | 5225703 | - | CCA CCGTCCGTACACTGGATTAT | |
| Cas21aChCBF1_018 | ChCBF2 | 4 | 6514830 | 6514853 | + | TTTC GTGAGACTCGTCACCCAATT |
| ChCBF1 | 7 | 17908848 | 17908871 | - | AATTGGGTGACGAGTCTCAC GAAA | |
| ChCBF3 | 8 | 13812351 | 13812374 | - | AATCGGATGACGTGTCTCAC GAAA | |
| Cas21aChCBF1_029 | ChCBF2 | 4 | 6515260 | 6515283 | + | TTTT GCCGCCTCCGTCCGTACAAT |
| ChCBF1 | 7 | 17908391 | 17908414 | - | ATTGTACGGACGGAGGCGGC AAAA | |
| Cas21aChCBF1_030 | ChCBF2 | 4 | 6515261 | 6515284 | + | TTTG CCGCCTCCGTCCGTACAATG |
| ChCBF1 | 7 | 17908390 | 17908413 | - | CATTGTACGGACGGAGGCGG CAAA | |