Table 2. Multiple targets Cas12a candidates for AtCBF 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 (TAIR v10) and whether it corresponds to a CBF gene. In alignments, black boxes mark mismatches and a space separates PAM (TTTN) from sequence. Differences in the “N” position in the PAM are not marked.
| Candidate ID | Targets + Offtargets (L0 = 4, L1 = 3) | |||||
|---|---|---|---|---|---|---|
| A. thaliana | Gene | chrom | start | end | sense | sequence |
| Cas12aAtCBF1_011 | AtCBF2 | 4 | 13015814 | 13015837 | - | GCTGCCATCTCAGCGGTTTG GAAA |
| AtCBF1 | 4 | 13022299 | 13022322 | - | GCTGCCATCTCAGCGGTTTG GAAA | |
| Cas12aAtCBF1_012 | AtCBF2 | 4 | 13015827 | 13015850 | - | CGGTTTGGAAAGTCCCGAGC CAAA |
| AtCBF1 | 4 | 13022312 | 13022335 | - | CGGTTTGGAAAGTCCCGAGC CAAA | |
| ---- | 1 | 27242286 | 27242310 | + | TTTG GCTCGGGACTTTCAACACAG | |
| ---- | 3 | 8296023 | 8296047 | + | TTTG GCTCGGGACGTTCGAAAGCG | |
| ---- | 5 | 17806910 | 17806934 | + | TTTG GCTCGGGACATTCGACACGG | |
| ---- | 5 | 21618544 | 21618567 | - | CCGTCTCAAAAGTCCCGAGC CAAA | |
| ---- | 4 | 7932903 | 7932927 | + | TTTG GCTCGGCACTTTTGAAACCG | |
| ---- | 4 | 10190722 | 10190745 | - | CAGTTTGGAACGTTCCGAGC CAAA | |
| AtCBF3 | 4 | 13018744 | 13018767 | - | CGGTTTGAAATGTTCCGAGC CAAA | |
| Cas12aAtCBF1_014 | AtCBF2 | 4 | 13015902 | 13015925 | - | TTCTTTGACGAACTCCTCTG TAAA |
| AtCBF1 | 4 | 13022387 | 13022410 | - | TTCTTTGACGAACTCCTCTG TAAA | |
| AtCBF4 | 5 | 21117594 | 21117617 | - | TCCTCTGACGAACTCCTCTG TAAA | |
| Cas12aAtCBF1_015 | AtCBF2 | 4 | 13015924 | 13015947 | - | AATTGGGTGACGAGTCTCAC GAAA |
| AtCBF1 | 4 | 13022409 | 13022432 | - | AATTGGGTGACGAGTCTCAC GAAA | |
| AtCBF3 | 4 | 13018841 | 13018864 | - | TATTGGGTGACGAGTCTCAC GAAA | |
| AtCBF4 | 5 | 21117616 | 21117639 | - | AATCGGATGACGTGTCTCAC GAAA | |
| Cas12aAtCBF1_017 | AtCBF2 | 4 | 13016031 | 13016054 | - | AATCGGAGCCAAACATTTCA GAAA |
| AtCBF3 | 4 | 13018948 | 13018971 | - | AATCGGAGCCAAACATTTCA GAAA | |
| AtCBF1 | 4 | 13022507 | 13022530 | - | AATCGGAGCCAAACATTTCA GAAA | |
| ---- | 1 | 8279033 | 8279056 | - | AATCAGAGCCTAACACTTCA AAAA | |
| ---- | 3 | 9399469 | 9399493 | + | TTTA TGAAGTGTTTGGTTCCTATT | |
| Cas12aAtCBF1_018 | AtCBF2 | 4 | 13016032 | 13016055 | - | ATCGGAGCCAAACATTTCAG AAAA |
| AtCBF3 | 4 | 13018949 | 13018972 | - | ATCGGAGCCAAACATTTCAG AAAA | |
| AtCBF1 | 4 | 13022508 | 13022531 | - | ATCGGAGCCAAACATTTCAG AAAA | |
| ---- | 1 | 9505057 | 9505081 | + | TTTG CTGAAATGGTTGCCTCTAAT | |
| Cas12aAtCBF1_019 | AtCBF3 | 4 | 13018950 | 13018973 | - | TCGGAGCCAAACATTTCAGA AAAA |
| AtCBF1 | 4 | 13022509 | 13022532 | - | TCGGAGCCAAACATTTCAGA AAAA | |
| Cas12aAtCBF1_024 | AtCBF2 | 4 | 13015842 | 13015865 | + | TTTG GAAAGTCCCGAGCCAAATCC |
| AtCBF1 | 4 | 13022327 | 13022350 | + | TTTG GAAAGTCCCGAGCCAAATCC | |
| ---- | 3 | 8296020 | 8296043 | - | GGGTTTGGCTCGGGACGTTC GAAA | |
| Cas12aAtCBF1_028 | AtCBF2 | 4 | 13015913 | 13015936 | + | TTTC TTTGACGAACTCCTCTGTAA |
| AtCBF1 | 4 | 13022398 | 13022421 | + | TTTC TTTGACGAACTCCTCTGTAA | |
| ---- | 5 | 16311156 | 16311179 | + | TTTT TTTGACGAATTTCTCTGTGG | |
| Cas12aAtCBF1_029 | AtCBF2 | 4 | 13015917 | 13015940 | + | TTTG ACGAACTCCTCTGTAAATTG |
| AtCBF1 | 4 | 13022402 | 13022425 | + | TTTG ACGAACTCCTCTGTAAATTG | |