Troubleshooting Off-Target Effects in CRISPR-Cas9 Experiments

[cevolution]

Hi all,

I’m encountering some unexpected off-target effects in my CRISPR-Cas9 experiments, even after using high-specificity sgRNAs and careful design through various online tools. While the on-target edits are fine, off-target mutations are showing up in regions I didn't expect.
slice masters
Has anyone else experienced this, and if so, what strategies did you use to minimize off-target effects? Are there particular design tools, reagents, or techniques you would recommend to reduce this issue?

Appreciate any insights or suggestions!