Table 1. 5 different sgRNA designations.
| sgRNA | Sequence (5′–3′) | Target sequence length | |
|---|---|---|---|
| sROSA26-sgRNA1 | target sequence | ggaggcgatgacgagatcgc ggg | 20 + PAM |
| sROSA26-sgRNA1-Up | cacc ggaggcgatgacgagatcgc | ||
| sROSA26-sgRNA1-Dn | aaac gcgatctcgtcatcgcctcc | ||
| sROSA26-sgRNA2 | target sequence | ggggagggagggattcttct agg | 20 + PAM |
| sROSA26-sgRNA2-Up | cacc ggggagggagggattcttct | ||
| sROSA26-sgRNA2-Dn | aaac agaagaatccctccctcccc | ||
| sROSA26-sgRNA3 | target sequence | gacgagatcgcgggggaggg agg | 20 + PAM |
| sROSA26-sgRNA3-Up | cacc ggacgagatcgcgggggaggg | ||
| sROSA26-sgRNA3-Dn | aaac ccctcccccgcgatctcgtcc | ||
| sROSA26-sgRNA4 | target sequence | gaggcgatgacgagatcgcg ggg | 20 + PAM |
| sROSA26-sgRNA4-Up | cacc ggaggcgatgacgagatcgcg | ||
| sROSA26-sgRNA4-Dn | aaac cgcgatctcgtcatcgcctcc | ||
| sROSA26-sgRNA5 | target sequence | gtcgagtctctcctcgatta tgg | 20 + PAM |
| sROSA26-sgRNA5-Up | cacc ggtcgagtctctcctcgatta | ||
| sROSA26-sgRNA5-Dn | aaac taatcgaggagagactcgacc |
5 guide RNAs (sgRNAs) were designed to target the sequence of the sRosa26 locus. The activity of the CRISPR/Cas9 was assessed using a UCA kit. All 5 sgRNAs efficiently guided Cas9 for genome editing.