Table 1.
Potential off-target sites for the gRNA used to generate Slc1a4E256K knock-in mice
| Target sequence1 | Score2 | Coordinates3 | Overlap | PCR primers |
|---|---|---|---|---|
| cTTCTTCAATTCCcTCAATGTGA | 6.8 | chr5:127662741-127662764− | intergenic | F: TTCCAGCTAGGGTGAAATGG R: TTCTGGTTTTCATGGGTGGT |
| cTTCTTCAATTCCcTCAATGTGA | 6.8 | chr18:41975999-41976022+ | Gm50410, intron 1 | F: CAACCATTGGACTGAGAGCA R: CACCCACCCACTCTTGTCTT |
| cTTCTcCAAcTCCTcCAATGTGG | 4.11 | chr5:56272235-56272258+ | intergenic | F: CAGGTGTGCCTGACAGAATG R: TGCCTTCTACCCTACGAAGC |
| cTTCTcCAAcTCCTcCAATGGAG | 3.7 | chr15:40158983-40159006+ | intergenic | ND (no sequence conservation) |
| tTTCTctAATTCCTTCAATGGGG | 3.28 | chr6:11131807-11131830− | intergenic | ND (conservation only with rat) |
| cTcCTcCAAcTCCTcCAATGGGG | 2.8 | chrX:141427255-141427278− | intergenic | ND (no sequence conservation) |
| tacCcTCAATTCCTcCAATGTGA | 2.61 | chr15:103060701-103060724− | intergenic | F: ACCTCCAGTGGCCAGAATTA R: AGGACACACCCAGTGACCTC |
| tTTCTTCAAcTCCTcCAATtAGG | 2.35 | chr19:39573727-39573750− | intergenic | ND (no sequence conservation) |
| tTTCTTCAAcTCCaTCAATGTGA | 2.17 | chr1:190941062-190941085− | Atf3, intron 1 | F: AAACAAGTCGCAACCCTGAC R: GTTCTCAGACCTGCCACTCC |
| gaTCTTCAATaCCTTCAATtGGG | 0.4 | chr2:62075707-62075730+ | Slc4a10, intron 7 | F: CAGCATTATCATTGCCGAAA R: TGATGGACCACAGCAACTCT |
| gTgCTTCAAaTCCTTCAATa | 0.86 | chr4:101010703-101010722+ | Jak1, 5’ UTR/promoter | F: GTGATCCACCAAGCCAACTT R: TTCGGTTGTTCCAGAAGGAG |
1
Lowercase indicates mismatches to Slc1a4 target sequence (ATTCTTCAATTCCTTCAATGAGG)
2
Scores for first 10 listed is from CRISPRoff, score for Jak1 5’UTR is from Benchling
3
+ or − after the genomic coordinates indicates sense or antisense strand