Table 2. List of the sequence for the CRISPR design.
| Strain | Genotype | Description | Sequence name | Sequence 5′−3′ | PAM | |
|---|---|---|---|---|---|---|
| AG406 | pezo-1 (av144) IV | Deletion of exons 1–13 and introns of pezo-1 | crRNA N-terminus | ACACAGCAACAACAGAATGA | CGG | |
| crRNA C-terminus | TGGGGGTGTTGCAGTGGCTA | AGG | ||||
| Repair template | atctgaatcggtggtcgtaacacagcaacaacagagtttgacacattttccgttgagacttgaaaaatag | |||||
| Genotyping F1 | GCGGTAAATCTGAATCGGTGG | |||||
| Genotyping R1 | TTGGAAAAGCAGGCACAACC | |||||
| Genotyping internal |
CGATCCAGCGTGGATGAACT | |||||
| AG416 | pezo-1 (av149) IV | Deletion of exons 27–33 and introns of pezo-1 | crRNA N-terminus | CGGTGGCAGCGTACATTATC | TGG | |
| crRNA C-terminus | CACCAGCGACACTCATCGAA | TGG | ||||
| Repair template | tccagtctcccatatttattttttttctgttccagTAGATAAGTAAGAGCAAAAAGAAGCAAGAATAA | |||||
| Genotyping F1 | AATCTGACTTGTGCCCTCCG | |||||
| Genotyping R1 | AATCAGGCGAGCAGTGAGAG | |||||
| Genotyping internal |
TCCACAGTCAATTCCTGCGT | |||||
| AG404 | pezo-1(av142 [mScarlet::pezo-1]) IV | Knock in mScarlet at N-terminus of pezo-1, mScarlet was amplified from plasmid pMS050 | crRNA | ACACAGCAACAACAGAATGA | CGG | |
| Repair template F1 | tgaatcggtggtcgtaacacagcaacaacagaATG CTTGTAGAGCTCGTCCATTCC (mScarlet) | |||||
| Repair template R1 |
AATTTGACGACGCACGATTTTAAAAGCGGCGGGACTGT
CTTGTAGAGCTCGTCCATTCC (mScarlet) |
|||||
| AG408 | pezo-1(av146 [gfp::pezo-1]) IV | Knock in GFP at N-terminus of pezo-1, GFP was amplified from plasmid pDD282 | crRNA | ACACAGCAACAACAGAATGA | CGG | |
| Repair template F1 | tgaatcggtggtcgtaacacagcaacaacagaATG agtaaaggagaagaattgttc (GFP) | |||||
| Repair template R1 |
AATTTGACGACGCACGATTTTAAAAGCGGCGGGACTGT
CTTGTAGAGCTCGTCCATTC (GFP) |
|||||
| AG483 | pezo-1(av182 [pezo-1::mScarlet]) IV. | Knock in mScarlet at C-terminus of pezo-1, mScarlet was amplified from plasmid pMS050 | NEST1 crRNA | CACCAGCGACACTCATCGAA | TGG | |
| Repair template |
AATATTCCTGTTCCGATCACCAGCGACACTCATCGAATGGACTCGTATGAGTAAGAAAAAACAGGAG
GTCTCCAAGGGAGAGGCCGTCATCAAGGAGTTCATGCGTTTCAAGGTCCAAGCGCTCCGAGGGACGTCACTCCACCGGAGGAATGGACGAGCTCTACAAGTAAatttaaatatttcactgtcaaatattctgcga (mScarlet) |
|||||
| Genotyping F1 | TGGTTCGAGAAGCGAAGGAC | |||||
| Genotyping R1 | aatcaggcgagcagtgagag | |||||
| NEST2 crRNA | TTCAAGGTCCAAGCGCTCCG | AGG | ||||
| Repair template F1 | GCCGTCATCAAGGAGTTCATGCGTTTCAAGGTCCACATGGAGGGATCCATGAACG | |||||
| Repair template R1 | TAGAGCTCGTCCATTCCTCCGGTGGAGTGACGTCCTTCTGAACGCTCGTATTGCTCGACGACGGTG | |||||
| AG487 | pezo-1(av190 [pezo-1::degron]) IV | Knock in Degron sequence at C-terminus of pezo-1, Degron was amplified from plasmid pK0132 | crRNA | CACCAGCGACACTCATCGAA | TGG | |
| Repair template F1 | AATATTCCTGTTCCGATCACCAGCGACACTCATCGAATGGACTCGTATGAGTAAGAAAAAACAGGAGggagcatcgggagcctcaggagcatcg (linker)GACTACAAAGACCATGACGGTG (Degron) | |||||
| Repair template R1 | tcgcagaatatttgacagtgaaatatttaaatTTACTTCACGAACGCCGCC (Degron) | |||||
| AG437 | pezo-1(av165[R2405P]) IV | Generate a point mutation R2405P in pezo-1 | crRNA | CTATTTGGTTCGAGAAGCGA | AGG | |
| Repair template | CATCTTCTCAAAATTTGTCTCGACATCTATTTGGTACCAGAAGCGAAAGACTTCATGTTGGAGCAGgtaattatttagtttta | |||||
| AG570 | pezo-1(av240) IV | Deletion of full length of pezo-1 | crRNA1 | ACACAGCAACAACAGAATGA | CGG | |
| crRNA2 | CACCAGCGACACTCATCGAA | TGG | ||||
| Repair template | ctgaatcggtggtcgtaacacagcaacaacagaATGTAGATAAGTAAGAGCAAAAAGAAGCAAGAATAAatttaaatatttc | |||||
| AG571 | pezo-1(av242) IV | Deletion of exons 27–33 and introns of pezo-1 in fem-1(hc17) | crRNA1 | CGGTGGCAGCGTACATTATC | TGG | |
| crRNA2 | CACCAGCGACACTCATCGAA | TGG | ||||
| Repair template | tccagtctcccatatttattttttttctgttccagTAGATAAGTAAGAGCAAAAAGAAGCAAGAATAA | |||||
| AG582 | pezo-1(av241) IV | Knock in Degron sequence at C-terminus of pezo-1 in AG404,Degron was amplified from plasmid pK0132 | crRNA | CACCAGCGACACTCATCGAA | TGG | |
| Repair template F1 | AATATTCCTGTTCCGATCACCAGCGACACTCATCGAATGGACTCGTATGAGTAAGAAAAAACAGGAGggagcatcgggagcctcaggagcatcg (linker)GACTACAAAGACCATGACGGTG (Degron) | |||||
| Repair template R1 | tcgcagaatatttgacagtgaaatatttaaatTTACTTCACGAACGCCGCC (Degron) | |||||
| PS8111 | pezo-1(sy1199) IV | Knock in a stop cassette at C-terminus of pezo-1 | crRNA | CCAGAAGCTCGTAAGCCAGG | AGG | |
| Repair template | cttatcgctgtttctgaaccagaagctcgtaagccGGGAAGTTTGTCCAGAGCAGAGGTGACTAAGTGATAAgctagcaggaggcactgaagaaacggatggtgatgaag | |||||
| Genotyping F1 | GACAGGACTTTCCCGCCAACTTAA | |||||
| Genotyping R1 | ATCATTCGCCGATTGCACAAGTTG | |||||
| PS8546 | pezo-1(sy1398) IV | Deletion of the first exon of pezo-1 isoforms i and j | crRNA1 | gagaacttgaattcaatgg | AGG | |
| crRNA2 | aagcttcttccgtctccgg | CGG | ||||
| crRNA3 | gcagtatttgaccaactgg | TGG | ||||
| crRNA4 | ataaaacaaggcaaccagg | GGG | ||||
| Genotyping F1 | CTCTCGCCTATCCACTTGAGCTTA | |||||
| Genotyping R1 | GGAAACAATTGAGCCGAGAATGGA | |||||
Note: Capital letters represent the ORF or exon sequence, small letters indicate the intron sequence. Bolded letters indicate the optimized bases needed for the CRISPR design.