Table 2.
pegRNA sequences (nucleotides in small letters represent the intended edit).
| Names | Spacer sequences | PBS sequences | RTT sequences | sgRNA sequences for PE3 | sgRNA sequences for PE3b |
|---|---|---|---|---|---|
| (A) Substitution at the splice acceptor site of exon 51 (nucleotides in small letters are intended modification) | |||||
| PegRNA1 | ACCAGAGTAACAGTCTGAGT | CAGACTGTTACTC | TTTaaCTCCTACT | TGACTTATTGTTATTGAAAT,AATGCCATCTTCCTTGATGT | GACTCCTACTCAGACTGTTACTC |
| PegRNA2 | ACCAGAGTAACAGTCTGAGT | CAGACTGTTACTC | TATTTTaaCTCCTACT | TGACTTATTGTTATTGAAAT, AATGCCATCTTCCTTGATGT | GACTCCTACTCAGACTGTTACTC |
| PegRNA3 | ACCAGAGTAACAGTCTGAGT | CAGACTGTTACTCTG | aaCTCCTACT | TGACTTATTGTTATTGAAAT, AATGCCATCTTCCTTGATGT | GACTCCTACTCAGACTGTTACTC |
| PegRNA4 | ACCAGAGTAACAGTCTGAGT | CAGACTGTTACTCTG | TTTaaCTCCTACT | TGACTTATTGTTATTGAAAT, AATGCCATCTTCCTTGATGT | GACTCCTACTCAGACTGTTACTC |
| PegRNA5 | ACCAGAGTAACAGTCTGAGT | CAGACTGTTACTCTG | TATTTTaaCTCCTACT | TGACTTATTGTTATTGAAAT, AATGCCATCTTCCTTGATGT | GACTCCTACTCAGACTGTTACTC |
| (B) Insertion of A at the end of exon 51 | |||||
| PegRNA1 | CGAGATGATCATCAAGCAGA | GCTTGATGATCATCTCGT | TTTCTCATACtCTTCT | CATCATTAAATTACAATCTA | |
| PegRNA2 | CGAGATGATCATCAAGCAGA | GCTTGATGATCATCT | ATTTTTTCTCATACtCTTCT | CATCATTAAATTACAATCTA | |
| PegRNA3 | CGAGATGATCATCAAGCAGA | GCTTGATGATCATCT | CCAACTTTTATCATTTTTTCTCATACtCTTCT | CATCATTAAATTACAATCTA | |
| PegRNA4 | CGAGATGATCATCAAGCAGA | GCTTGATGATCATCT | TTTCTCATAtCaTTCT | CATCATTAAATTACAATCTA | |
| PegRNA5 | CGAGATGATCATCAAGCAGA | GCTTGATGATCATCT | TTTCTCATAtCaTTCT | CATCATTAAATTACAATCTA | |
| PegRNA6 | CGAGATGATCATCAAGCAGA | GCTTGATGATCATCT | TTTCTCATAtaaTTCT | CATCATTAAATTACAATCTA | |
| (C) pegRNAs optimization to increase the editing efficiency | |||||
| SubSpD51−1 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCTTTTCTCATtgCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−2 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCTCTGCCAACTTTTATCATTTTTTCTCATtgCTTtTGC | CATCATTAAATTACAATCTA | |
| SubSpD51−3 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCAAAAAAGAAAAACTTCTGCCAACTTTTATCATTTTTTCTCATtgCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−4 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCTTTTCTCATggCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−5 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCTCTGCCAACTTTTATCATTTTTTCTCATggCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−6 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCAAAAAAGAAAAACTTCTGCCAACTTTTATCATTTTTTCTCATggCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−7 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCTTTTCTCATcgCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−8 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCTCTGCCAACTTTTATCATTTTTTCTCATcgCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−9 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | GTGCAAAAAAGAAAAACTTCTGCCAACTTTTATCATTTTTTCTCATcgCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−10 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATcgCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−11 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATcgtTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−12 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATggtTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−13 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATcaCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−14 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATcatTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−15 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | CATcgCTTtT | CATCATTAAATTACAATCTA | |
| SubSpD51−16 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | CATggCTTtT | CATCATTAAATTACAATCTA | |
| (D) Insertion of nucleotides at the exon 51 splice donor site | |||||
| InsSpD51+6–1 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATAtCCTTCT | CATCATTAAATTACAATCTA | |
| InsSpD51+6–2 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATAgCCTTCT | CATCATTAAATTACAATCTA | |
| InsSpD51+6–3 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATAtgCCTTCT | CATCATTAAATTACAATCTA | |
| InsSpD51+6–4 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATAtgtCCTTCT | CATCATTAAATTACAATCTA | |
| InsSpD51+6–5 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATAgggCCTTCT | CATCATTAAATTACAATCTA | |
| InsSpD51+6–6 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TTTTCTCATAtttCCTTCT | CATCATTAAATTACAATCTA | |
| (E) Deletion of nucleotides in exon 51 splice donor site | |||||
| DelSpD51+6–1 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | ATTTTTTCTCATCTTCT | CATCATTAAATTACAATCTA | |
| DelSpD51+6–2 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | TCATTTTTTCTCCTTCT | CATCATTAAATTACAATCTA | |
| DelSpD51+6–3 | CGAGATGATCATCAAGCAGA | GCTTGATGATCAT | ATCATTTTTTCCTTCT | CATCATTAAATTACAATCTA | |
| (F) Substitution at +6 in the splice donor site of exon 53 | |||||
| SubSpD53+6–1 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTT | CTAggaTTGG | CATCATTAAATTACAATCTA | |
| SubSpD53+6–2 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTT | ATACTAggaTTGG | CATCATTAAATTACAATCTA | |
| SubSpD53+6–3 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTT | CTAcgaTTcG | CATCATTAAATTACAATCTA | |
| SubSpD53+6–4 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTT | TTGATACTAggaTTGG | CATCATTAAATTACAATCTA | |
| SubSpD53+6–5 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTT | TTGATACTAcgaTTGG | CATCATTAAATTACAATCTA | |
| SubSpD53+6–6 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTT | TTGATACTAggaTTcG | CATCATTAAATTACAATCTA | |
| SubSpD53+6–7 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTT | TTGATACTAcgaTTcG | CATCATTAAATTACAATCTA | |
| (G) Substitution at +1 in the splice donor site of exon 53 | |||||
| SubSpD53+1–1 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGA | GAAACgAAac | CCAGAGCCAAGCTTGAGTCA | |
| SubSpD53+1–2 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGA | GAAACaAAac | CCAGAGCCAAGCTTGAGTCA | |
| SubSpD53+1–3 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGA | ACAGAAACgAAac | CCAGAGCCAAGCTTGAGTCA | |
| SubSpD53+1–4 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGA | ACAGAAACaAAac | CCAGAGCCAAGCTTGAGTCA | |
| SubSpD53+1–5 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGA | ATCACAGAAACgAAac | CCAGAGCCAAGCTTGAGTCA | |
| SubSpD53+1–6 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGA | ATCACAGAAACaAAac | CCAGAGCCAAGCTTGAGTCA | |
| (H) Insertion of nucleotides at the splice donor of exon 53 | |||||
| InsSpD53+6–1 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | ATACTAgggggaTTcG | CATCATTAAATTACAATCTA | |
| InsSpD53+6–2 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | TTGATACTAgggggaTTcG | CATCATTAAATTACAATCTA | |
| InsSpD53+6–3 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | TACTAggccgggaTTcG | CATCATTAAATTACAATCTA | |
| InsSpD53+6–4 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | GATACTAggccgggaTTcG | CATCATTAAATTACAATCTA | |
| InsSpD53+1–1 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGA | ACgAAacccc | CCAGAGCCAAGCTTGAGTCA | |
| InsSpD53+1–2 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGATA | GAAACgAAacccc | CCAGAGCCAAGCTTGAGTCA | |
| InsSpD53+1–3 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGATA | ACAGAAACgAAacccc | CCAGAGCCAAGCTTGAGTCA | |
| InsSpD53+1–4 | GGTATCTTTGATACTAACCT | TTAGTATCAAAGATA | AGAAACgAAaccggcc | CCAGAGCCAAGCTTGAGTCA | |
| (I) Deletion at +6 of nucleotides at the splice donor of exon 53 | |||||
| DelSpD53+6–1 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | TTGATACTAaTTGG | CATCATTAAATTACAATCTA | |
| DelSpD53+6–2 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | ctTTGATACTAaGG | CATCATTAAATTACAATCTA | |
| DelSpD53+6–3 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | atctttgatactGG | CATCATTAAATTACAATCTA | |
| DelSpD53+6–4 | AAAGAAAATCACAGAAACCA | TTTCTGTGATTTTCT | TTGATACTACTTGG | CATCATTAAATTACAATCTA | |
| DelSpD53+1–1 | AAAGAAAATCACAGAAACCA | TTAGTATCAAAGATA | ATCACAGAAACaAAa | CCAGAGCCAAGCTTGAGTCA | |