Table 5.
Size of the expected amplicons and additional information for CRISPR editing methods.
| Portion to KO | gRNA pairs used | Guides sequence | Expected fragment size if portion KO (bp) | Expected fragment size if portion is NOT KO (bp) | F primer sequence | R primer sequence | |
|---|---|---|---|---|---|---|---|
| CRE3-1 | 1 | 3’ guide | TCAAATCATGACACCCTGAA | 659 | 1432 | GAAAGTCAGGGCCAGTTGAC | AAGGTTGTCGGGATAAGCGG |
| 5’ guide | CTGGGCAAGGACCATTCCCA | ||||||
| CRE3-2 | 2 | 3’ guide | TCAAATCATGACACCCTGAA | 836 | 1653 | GAAAGTCAGGGCCAGTTGAC | TCTTGGATCTCCAGCCTGTC |
| 5’ guide | TGTCAGATACAAATCACACC | ||||||
| CRE3-3 | 3 | 3’ guide | TCAAATCATGACACCCTGAA | 608 | 1690 | GAAAGTCAGGGCCAGTTGAC | GGAGAACTGCAAGGTTCAGGA |
| 5’ guide | TAAGCGGTGAAACATTACCA | ||||||
| CRE6-1 | 4 | 3’ guide | ACATCAATTATATTAAACGT | 608 | 1773 | CGTCACGTTCTATGAGAGTCC | ACTCCTAGCGACCCATCACT |
| 5’ guide | caagtagctaggactacagg | ||||||
| CRE6-2 | 5 | 3’ guide | TTCCTTTAGCaaaaaaaaCG | 642 | 1677 | AGTCCAGGCATCTGAGGTTC | ACTCCTAGCGACCCATCACT |
| 5’ guide | caagtagctaggactacagg | ||||||
| CRE 3 to 6 | 6 | 3’ guide | TCAAATCATGACACCCTGAA | 637 | Portion 1a: 1432 | GAAAGTCAGGGCCAGTTGAC | AAGGTTGTCGGGATAAGCGG |
| 5’ guide | caagtagctaggactacagg | Portion 2a: 1773 | CGTCACGTTCTATGAGAGTCC | ACTCCTAGCGACCCATCACT | |||
| For fragment cuta | GAAAGTCAGGGCCAGTTGAC | ACTCCTAGCGACCCATCACT | |||||
| OR2W5 | 7 | 3’ guide | GACAACCAGGAGGACGCACT | 500 | 700 | 5′-TCGGCCTGGACTGGAGAAAA-3′ | 5′-GAGACCACTGTGAGGTGAGA-3′ |
| 5’ guide | CTCCCGGTGTGGACGTCGCA |
aGiven that the region from CRE3 to CRE6 was too long to amplify with classic PCR, only a portion of the region was sequenced using the indicated primers, starting from one side (primers F and R1) or the other side (portion 1) or the other side (portion 2), or the full region in the case same has been KO (for fragment cut), leading to the following expected fragments sizes.