Table 1 NmeCas9 or SpyCas9 guide and target sequences used in this study. NTS, NmeCas9 target site; STS, SpyCas9 target site. The sgRNA spacer sequences (5′➔3′) are shown with their canonical lengths, and with a 5′-terminal G residue; non-canonical lengths are described in the text and figures. Target site sequences are also 5′➔3′ and correspond to the DNA strand that is non-complementary to the sgRNA, with PAM sequences underlined. The names of sites that showed at least 3% editing with NmeCas9 are indicated in bold
From: NmeCas9 is an intrinsically high-fidelity genome-editing platform
NmeCas9 Site | Gene or locus | Spacer Sequence | Target site, with PAM |
|---|---|---|---|
Ps9 | N.A. (reporter) | GCAUCCUCAGAUUUAGUAUUCAGA | ATGACATCCTCAGATTTAGTATTCAGA ATATGATT TTC |
Ps24 | N.A. (reporter) | GUCGCGCAACAAAACCCCGCUACU | GTCGTCGCGCAACAAAACCCCGCTACT CGGCGATT TTC |
Ps25 | N.A. (reporter) | GCAUGCGCGGCGCAUUACCUUUAC | TCCTCATGCGCGGCGCATTACCTTTAC GCCGGATT GCC |
NTS1B | SLC9A9 | GGGCAUCAUGAUUUUGAACUCCCU | CCTTGGCATCATGATTTTGAACTCCCT ATGTGATT CTA |
NTS1C | SLC9A9 | GUGGUCUGGGGUACAGCCUUGGCA | TACTUGGTCTGGGGTACAGCCTTGGCA TCATGATT TTG |
NTS1C-OT1 | PHKG2 | GCGGUGUGAGGUACAGCCUUGGCA | TAATCGGTGTGAGGTACAGCCTTGGCA TCAGGATT TCT |
NTS3 | AL158836 | GAUGCUCAGAAAGAGGAAGCUGGU | GGGGATGCTCAGAAAGAGGAAGCTGGT TTATGATT GGA |
NTS4B | FLJ00328 | GGACAGGAGUCGCCAGAGGCCGGU | GCAGGACAGGAGTCGCCAGAGGCCGGT GGTGGATT TCC |
NTS4C | FLJ00328 | GGGGCUGGCUCCACGUCGCGCCGC | TGCGGGGCTGGCTCCACGTCGCGCCGC GGCGGTTT GGG |
NTS5 | AF064860 | GAAACAGACUCGCAAGACUUCAGA | GACAAAACAGACTCGCAAGACTTCAGA TACAGATT CCA |
NTS7 | LOC100505797 | GAGGGAGAGAGGUGAGCGGAUGAA | GCAAAGGGAGAGAGGTGAGCGGATGAA GGGAGATT GGT |
NTS8 | ESPN | GGACGCAAUUCCAGAGGUGAUGGG | CGGCGACGCAATTCCAGAGGTGATGGG GAGTGATT GTC |
NTS9 | ZNHIT2 | GGCGCUGUGUUUUCGCAAAGCUUC | CGGCGCGCTGTGTTTTCGCAAAGCTTC CGAGGATT CTC |
NTS10 | HHLA1 | GCAGCCAAGUUUGAGAACUGCUGU | TGTGCAGCCAAGTTTGAGAACTGCTGT TACAGATT TCC |
NTS11 | SMARCB1 | GUUCCAGUUGGGAAGGGCCAGUGC | TAGATTCCAGTTGGGAAGGGCCAGTGC CTCCGATT CCA |
NTS20 | CH251-396E2 | GAUACCAGUCAGAUUUGUGCAAUG | TCTAATACCAGTCAGATTTGTGCAATG TTGGGTTT GGA |
NTS21 | TNNC1 | GCCAGAGCUGCCGCCAGACAGUGA | CAGTCCAGAGCTGCCGCCAGACAGTGA TGCTGTCT TGG |
NTS22 | ANO1 | GUUUGGCUGGGCGCUCCUGAAACU | TGCTTTTGGCTGGGCGCTCCTGAAACT CTGGGCTT TCA |
NTS23 | LOC103240889 | GCGGCCCCGGGGCCGGGCCGCGGG | GCGACGGCCCCGGGGCCGGGCCGCGGG CGGAGACT GAC |
NTS24 | AC097382 | GCUGCCUCAUGGUGUGUGCUGAGG | GCTGCTGCCTCATGGTGTGTGCTGAGG CTCAGACA TCA |
NTS25 | AC193513 | GGUUUCUCAUCCUGUCUUCUGCCU | CCGCGTTTCTCATCCTGTCTTCTGCCT AGTGGATA TGT |
NTS26 | LOC105378512 | GUUCAAAAGUAGCGGGCGCUAGGC | GTACTTCAAAAGTAGCGGGCGCTAGGC GGGTGTTT CTG |
NTS27 | TIE1 | GUUCUCCAAGCCCUCGGACCUCGU | CGGCTTCTCCAAGCCCTCGGACCTCGT GGGCGTCT TCT |
NTS28 | AL138885 | GCAGGGGCAGGGGCGCAUCAGCUG | GCGGCAGGGGCAGGGGCGCATCAGCTG TCCGGCTT GGA |
NTS30 | NEK8 | GGGGCUCCGGAGCCCACCCCAGGA | CGCGGGGCTCCGGAGCCCACCCCAGGA CCAGGACT TAG |
NTS31 | POC1A | GUGGGAAGUGUAGCUCCACCUUCC | ATGTTGGGAAGTGTAGCTCCACCTTCC TCCAGACA TAG |
NTS32 | VEGFA | GCCCCGGCUCUGGCUAAAGAGGGA | CACACCCCGGCTCTGGCTAAAGAGGGA ATGGGCTT TGG |
NTS33 | VEGFA | GCGGGGAGAAGGCCAGGGGUCACU | GGAGCGGGGAGAAGGCCAGGGGTCACT CCAGGATT CCA |
NTS55 | CYBB | GCUGGAUUACUGUGUGGUAGAGGG | CTAGCTGGATTACTGTGTGGTAGAGGG AGGTGATT AGC |
NTS58 | AAVS1 | GUUUGCCUGGACACCCCGUUCUCC | TTTCTTTGCCTGGACACCCCGTTCTCC TGTGGATT CGG |
NTS59 | AAVS1 | GACCCCACAGUGGGGCCACUAGGG | CTCCACCCCACAGTGGGGCCACTAGGG ACAGGATT GGT |
NTS63 | AAVS1 | GAGUUAGAACUCAGGACCAACUUA | CCAAAGTTAGAACTCAGGACCAACTTA TTCTGATT TTG |
NTS64 | Rosa26 | GGCAGAUCACGAGGGAAGAGGGGG | AGTTGCAGATCACGAGGGAAGAGGGGG AAGGGATT CTC |
DTS1-NmeCas9 | KL | GCUGGCUGAGCCCAUUUUCGGCUC | CTGGCTGGCTGAGCCCATTTTCGGCTC TGGAGATT ATC |
DTS3-NmeCas9 | ARHGEF9 | GACUGAAGGCGAGGUCCGGGGCGG | ACTGACTGAAGGCGAGGTCCGGGGCGG AGGGGATT GGG |
DTS7-NmeCas9 | LSP1 | GGCUGGCACCCUCCAUGUACCCAG | GCCGGCTGGCACCCTCCATGTACCCAG GGGAGATT CCA |
DTS8-NmeCas9 | PCDHB4 | GUGAGUGAAGAUAUGAUAAGUUCU | CTGGTGAGTGAAGATATGATAAGTTCT GGGGGATT GTC |
SpyCas9 Site | Gene or locus | Spacer Sequence | Target site, with PAM |
STS60 | AAVS1 | GUUAAUGUGGCUCUGGUUCU | CCGGTTAATGTGGCTCTGGTTCT GGG TAC |
STS61 | AAVS1 | GUCCCCUCCACCCCACAGUG | TCTGTCCCCTCCACCCCACAGTG GGG CCA |
STS62 | AAVS1 | GGGGCCACUAGGGACAGGAU | AGTGGGGCCACTAGGGACAGGAT TGG TGA |
DTS1-SpyCas9 | KL | GCUGAGCCCAUUUUCGGCUC | CTGGCTGAGCCCATTTTCGGCTC TGG AGA |
DTS3-SpyCas9 | ARHGEF9 | GAAGGCGAGGUCCGGGGCGG | ACTGAAGGCGAGGTCCGGGGCGG AGG GGA |
DTS7-SpyCas9 | LSP1 | GGCACCCUCCAUGUACCCAG | GCTGGCACCCTCCATGTACCCAG GGG AGA |
DTS8-SpyCas9 | PCDHB4 | GUGAAGAUAUGAUAAGUUCU | TGAGTGAAGATATGATAAGTTCT GGG GGA |