Table 2.
Filtered features | Excluded value | Enrichment ratio for non-functional gRNA |
---|---|---|
gRNA folding (∆G) | < −8 kcal/mol | 15.8 |
Duplex binding (∆G) | < −22 kcal/mol | 3.5 |
GC content | >80 % | 30.7 |
UUU in the seed region | True | 10.5 |
Repetitive bases | True | 4.2 |
Position 19 | U | 2.6 |
Position 20 | C or U | 2.5 |
Free energy (∆G) was calculated by RNAfold for gRNA self-folding and by the nearest neighbor method for binding stability of gRNA–target duplex