Table 3.
Synthesized gRNA constructs. In gRNA construct sequences, bold sequences indicate the BsaI Type II restriction enzyme recognition site, while underlined sequences are the BsaI cutting site.
| Synthesized sequences | Sequence (5’-3’) |
|---|---|
| Self-cleaving elements | |
| HH | ATATTGCTGATGAGTCCGTGAGGACGAAACGAGTAAGCTCGTC |
| HDV | GGCCGGCATGGTCCCAGCCTCCTCGCTGGCGCCGGCTGGGCA ACATGCTTCGGCATGGCGAATGGGAC |
| tRNAGly | GCGCAAGTGGTTTAGTGGTAAAATTCATCGTTGCCATCGATGAGCCCCCGGTTCGATTCCGGG CTTGCGCA |
| tRNAAsp | TCCGTGATAGTTTAATGGTCAGAATGGGCGCTTGTCGCGTGCCAGATCGGGGTTCAATTCCCCGTCGCGGAG |
| Structural gRNA | GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAA AAAGTGGCACCGAGTCGGTGC |
| gRNA constructs | |
| tRNAGly- ADE2 target—tRNAAsp | CCCTTTAAGGTCTCGCAAAGCGCAAGTGGTTTAGTGGTAAAATTCATCGTTGCCATCGATGAGCCCC CGGTTCGATTCCGGGCTTGCGCACAATATGATAGCGTCGTCCAGTTTTAGAGCTAGAAATAGCAAGT TAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTCCGTGATAGT TTAATGGTCAGAATGGGCGCTTGTCGCGTGCCAGATCGGGGTTCAATTCCC CGTCGCGGAGACAGCGAGACCCCCTTTAA |
| tRNAGly- ADE2 target—HDV | CCCTTTAAGGTCTCGCAAAGCGCAAGTGGTTTAGTGGTAAAATTCATCGTTGCCATCGATGAGCCCCC GGTTCGATTCCGGGCTTGCGCACAATATGATAGCGTCGTCCAGTTTTAGAGCTAGAAATAGCAAGTTA AAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTGGCCGGCATGGTC CCAGCCTCCTCGCTGGCGCCGGCTGGGCAACATGCTTCGGCATGGCGAATGGGACACAGCGAGACCC CCTTTAA |
| HH—ADE2 target—tRNAAsp | CCCTTTAAGGTCTCGCAAAATATTGCTGATGAGTCCGTGAGGACGAAACGAGTAAGCTCGTCCAATAT GATAGCGTCGTCCAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGA AAAAGTGGCACCGAGTCGGTGCTTTTGCGCAAGTGGTTTAGTGGTAAAATTCATCGTTGCCATCGATG AGCCCCCGGTTCGATTCCGGGCTTGCGCAACAGCGAGACCCCCTTTAA |
| HH- ADE2 target—HDV | CCCTTTAAGGTCTCGCAAAATATTGCTGATGAGTCCGTGAGGACGAAACGAGTAAGCTCGTCCAATAT GATAGCGTCGTCCAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGA AAAAGTGGCACCGAGTCGGTGCTTTTGGCCGGCATGGTCCCAGCCTCCTCGCTGGCGCCGGCTGG GCAACATGCTTCGGCATGGCGAATGGGACACAGCGAGACCCCCTTTAA |
| tRNAGly- tRNAAsp-U1 | AAGGAAGCGTCTCACAAAGCGCAAGTGGTTTAGTGGTAAAATTCATCGTTGCCATCGATGAGCCCCC GGTTCGATTCCGGGCTTGCGCATGAGACCTCACACTTCACGGGTAGTATGGTAGGGTCTCCGTTTT AGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCG GTGCTTTTTCCGTGATAGTTTAATGGTCAGAATGGGCGCTTGTCGCGTGCCAGATCGGGGTTCA ATTCCCCGTCGCGGAGACAGCGAGACGCTTCCAGGG |