| Primer | Sequence 5′–3′ | Purpose |
|---|---|---|
| Kras_1F | 5′-TAATACGACTCACTATAGGAATATAAACTTGTGGTAGT-3′ | Generation of KRAS and NRAS gRNAs |
| Kras_1R | 5′-TTCTAGCTCTAAAACACTACCACAAGTTTATATTC-3′ | |
| Kras_2F | 5′-TAATACGACTCACTATAGAATGACTGAATATAAACTTG-3′ | |
| Kras_2R | 5′-TTCTAGCTCTAAAACCAAGTTTATATTCAGTCATT-3′ | |
| Nras_1F | 5′-TAATACGACTCACTATAGGACTGAGTACAAACTGGTGG-3′ | |
| Nras_1R | 5′-TTCTAGCTCTAAAACCCACCAGTTTGTACTCAGTC-3′ | |
| Nras_2F | 5′-TAATACGACTCACTATAGAATGACTGAGTACAAACTGG-3′ | |
| Nras_2R | 5′-TTCTAGCTCTAAAACCCAGTTTGTACTCAGTCATT-3′ | |
| KrasutrF1 | 5′-GCGTCGATGGAGGAGTTTG-3′ | Sequencing of KRAS and NRAS clones |
| KrasR2 | 5′-ACTCTTGCCTACGCCACCA-3′ | |
| NrasutrF1 | 5′-GGGGAGATCTTTGGAGACAGAA-3′ | |
| NrasR2 | 5′-TGGGTAAAGATGATCCGACAAGT-3′ | |
| grb2-RNA_T1 | 5′-ATCTTGTGGAAAGGACGAAACACCGGGTTAGACGTTCCGGTTCACGGGTTTTAGAGCTAGAAATAGCAAGTT-3′ | Generation of Grb2 gRNAs |
| grb2- gRNA_T2 | 5′-ATCTTGTGGAAAGGACG AAACACCGGGCTTAGACGTTCCGGTTCACGGTTTTAGAGCTAGAAATAGCAAGTT-3′ | |
| Grb-HA_f | 5′-TATCGATAAGCTTGATATCGAGAGGCAGTGTGTAGCCAG-3′ | Generation of Grb2-mNG donor |
| Grb-HA_r | 5′-CTCCTCCTAAGACGTTCCGGTTCACGGG-3′ | |
| Grb-Neon_f | 5′-CCGGAACGTCTTAGGAGGAGGAGGATCAG-3′ | |
| Grb-Neon_r | 5′-TTGACTCTTACTTGTACAGCTCGTCCATG-3′ | |
| Grb-3HA_f | 5′-GCTGTACAAGTAAGAGTCAAGAAGCAATTATTTAAAG-3′ | |
| Grb-3HA_r | 5′-CCACCGCGGTGGCGGCCGCTTCTCGAACTCCTGACCTTG-3′ | |
| mCh-F3 | 5′-ATGGTGAGCAAGGGCGAG-3′ | Generation of RAF1-mCh donor |
| mCH-R2 | 5′-CTTGTACAGCTCGTCCATGC-3′ | |
| mCh_F4 | 5′-CTTGCACGCTGACCACGTCCCCGAGGCTGCCTGTCTTCTTAGGAGGA-3′ | |
| 5′-GGAGGATCAGCAGCAGCAATGGTGAGCAAGGGCGAG-3′ | ||
| mCh_R3 | 5′-CTGAAGACAGGTGCAAAGTCAACTACTTGTACAGCTCGTCCATG-3′ | |
| CHC siRNA | 5′-GCAATGAGCTGTTTGAAGA-3′ | CHC knockdown |
| Primer | Sequence 5′–3′ | Purpose |
|---|---|---|
| Kras_1F | 5′-TAATACGACTCACTATAGGAATATAAACTTGTGGTAGT-3′ | Generation of KRAS and NRAS gRNAs |
| Kras_1R | 5′-TTCTAGCTCTAAAACACTACCACAAGTTTATATTC-3′ | |
| Kras_2F | 5′-TAATACGACTCACTATAGAATGACTGAATATAAACTTG-3′ | |
| Kras_2R | 5′-TTCTAGCTCTAAAACCAAGTTTATATTCAGTCATT-3′ | |
| Nras_1F | 5′-TAATACGACTCACTATAGGACTGAGTACAAACTGGTGG-3′ | |
| Nras_1R | 5′-TTCTAGCTCTAAAACCCACCAGTTTGTACTCAGTC-3′ | |
| Nras_2F | 5′-TAATACGACTCACTATAGAATGACTGAGTACAAACTGG-3′ | |
| Nras_2R | 5′-TTCTAGCTCTAAAACCCAGTTTGTACTCAGTCATT-3′ | |
| KrasutrF1 | 5′-GCGTCGATGGAGGAGTTTG-3′ | Sequencing of KRAS and NRAS clones |
| KrasR2 | 5′-ACTCTTGCCTACGCCACCA-3′ | |
| NrasutrF1 | 5′-GGGGAGATCTTTGGAGACAGAA-3′ | |
| NrasR2 | 5′-TGGGTAAAGATGATCCGACAAGT-3′ | |
| grb2-RNA_T1 | 5′-ATCTTGTGGAAAGGACGAAACACCGG | Generation of Grb2 gRNAs |
| grb2- gRNA_T2 | 5′-ATCTTGTGGAAAGGACG AAACACCGG | |
| Grb-HA_f | 5′-TATCGATAAGCTTGATATCGAGAGGCAGTGTGTAGCCAG-3′ | Generation of Grb2-mNG donor |
| Grb-HA_r | 5′-CTCCTCCTAAGACGTTCCGGTTCACGGG-3′ | |
| Grb-Neon_f | 5′-CCGGAACGTCTTAGGAGGAGGAGGATCAG-3′ | |
| Grb-Neon_r | 5′-TTGACTCTTACTTGTACAGCTCGTCCATG-3′ | |
| Grb-3HA_f | 5′-GCTGTACAAGTAAGAGTCAAGAAGCAATTATTTAAAG-3′ | |
| Grb-3HA_r | 5′-CCACCGCGGTGGCGGCCGCTTCTCGAACTCCTGACCTTG-3′ | |
| mCh-F3 | 5′-ATGGTGAGCAAGGGCGAG-3′ | Generation of RAF1-mCh donor |
| mCH-R2 | 5′-CTTGTACAGCTCGTCCATGC-3′ | |
| mCh_F4 | 5′-CTTGCACGCTGACCACGTCCCCGAGGCTGCCTGTCTTCTTAGGAGGA-3′ | |
| 5′-GGAGGATCAGCAGCAGCAATGGTGAGCAAGGGCGAG-3′ | ||
| mCh_R3 | 5′-CTGAAGACAGGTGCAAAGTCAACTACTTGTACAGCTCGTCCATG-3′ | |
| CHC siRNA | 5′-GCAATGAGCTGTTTGAAGA-3′ | CHC knockdown |
Underlined sequences correspond to target gRNA sequence.
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