Table 1 Targeted mutagenesis site, possible off-target sites, the respective protospacer adjacent motif (PAM) sequences, and forward (F) and reverse (R) oligonucleotide primers used for PCR amplification and DNA sequencing
Guide strand | PAM | Chromosome in human genome | Oligo primers used in sequencing (5′-3′) |
|---|---|---|---|
Targeted mutagenesis site | |||
TCTGTTGCTTCGAACTCCAG | TGG | 11 | GTTCGTTGCAACAAATTGAT (F) |
GTCAGTAACAACTTGTCCAA (R) | |||
Possible off-target sites | |||
TATG-TGCTTCGAGCTCCAG | AGG | 19 | CTTCTGTGCCTTACGTACCA (F) |
TGTACTGTAACCAAGTACGC (R) | |||
TCTCTTGCTTC-AACTCCAG | AGG | 3 | GTAGAGGCAAGGTCTTCCTG (F) |
TTACCAGCCTCCTTGGACCT (R) | |||
TCAGCTTGCTTCGAACTCCTG | GAG | 12 | TTAGCAATAACTAGTCCAAC (F) |
CAGAGGTCACTGTAGTGTGG (R) | |||