TABLE 2.
In silico scoring and in vivo editing of individual gRNAs. Efficiencies grated from highest to lowest, with one being the best scoring gRNA. Scores based on the gRNA individual score in the in silico prediction servers CHOPCHOP (http://chopchop.cbu.uib.no/), CRISPRater (https://crispr.cos.uni-heidelberg.de/), SSC(http://crispr.dfci.harvard.edu/SSC/). The in vivo score is based on editing efficiencies of single RNP/gRNA transformations as evidenced by IDAA (from two to eight biological replicates were performed with std errors indicated (see also Figures 4, 5).
| gRNA | CHOPCHOP | SSC | CRISPRater | In vivo | |
|---|---|---|---|---|---|
| GWD 5′ | gJ: TCAGTGGTAAGTACAGCATG | 1 | 1 | 1 | 1 |
| gK: AGGGAATAACTTGCTGTACC | 3 | 4 | 3 | 4 | |
| gL: GTTTCGAGGTAACAGGTTAA | 4 | 3 | 2 | 2 | |
| gM: GTACAGCAAGTTATTCCCTA | 2 | 2 | 2 | 3 | |
| GWD 3′ | gA: GGAGAGGAGGAAATTCCTGA | 2 | 5 | 1 | 5 |
| gB: TGTTCGAGCTAGAAATGGGA | 3 | 4 | 5 | 6 | |
| gC: GCTGACCTCCAAGCAAAGGA | 5 | 2 | 4 | 1 | |
| gD: ATTGGCTGACCTCCAAGCAA | 4 | 3 | 2 | 2 | |
| gE: TTTCTGTTCGAGCTAGAAAT | 6 | 6 | 3 | 4 | |
| sgI: CACAACGACAACATATCCAA | 1 | 1 | 6 | 3 | |
| DMR6 5′ | g43: TTTGAGGGAGAGTAGAGTGG | 2 | 1 | 1 | 1 |
| g44: GTGGCCTATCGGATTCGGGT | 1 | 2 | 2 | 3 | |
| g45: TGGAGAAATATGCTCCTGAA | 3 | 3 | 3 | 2 | |
| DMR6 3′ | g46: GAAGCCATAGCAGAGAGCCT | 2 | 2 | 1 | 2 |
| g47: GAATTTGGATCAGTATGGGC | 3 | 1 | 2 | 1 | |
| g48: ATCACCAAGATTAATGACAA | 1 | 3 | 3 | 3 |