When analyzing gRNAs, Benchling calculates two types of scores associated with off-target assessment:
- Aggregated off-target score
- Potential off-target site score
The meaning of the score depends on which off-target score you're referencing.
Aggregated off-target scores
Potential off-target site scores
|Each gRNA has one aggregated off-target score. The aggregation scoring approach is described in Hsu et al. 2013, but was also applied for the Doench method to provide a uniform set of scores.
|Benchling numbers and scores all potential off-target sites in selected genomes. This means each gRNA can have multiple potential off-target sites, each with their own score.
|How to view
|In the list of gRNA in the Design CRISPR tab
|In the pop-up window after clicking into each gRNA’s aggregate off-target score
|Higher aggregated off-target scores are generally better guides.
High scoring off-target sites indicate a high likelihood the gRNA will interact with that off-target site, which is generally not wanted. In general, A “good” gRNA with the highest aggregate off-target score should have either or both: