Cutting Once Is Harder Than It Sounds: Your Guide to Minimizing Off-Target Effects
Type
Blog Post
Published
March 31, 2026, 1:15 p.m.
... so minimal experimental testing is required.
So how can you do this? Testing all IFNs is completely unnecessary...are a matter of sensitivity. Therefore, the key question is not whether off-target effects exist, but what...simple: choose the least active SpCas9 variant that still efficiently cuts your on-target site.
If a nuclease...). However, only the least active variant which still retains on-target activity (highlighted by a bold...higher-activity nucleases, the weakest variant that is still active can achieve off-target–free editing for this...Figure 2).
Identify the least active variant that still shows the right amount of on-target editing.
This.... (In the example in Figure 3, the least active still working variant was B-HypaR, thus variants with ...