Improving the efficiency of high-fidelity Cas9 by enhancing PAM-distal interactions - PubMed
5 hours ago
- #Genome Editing
- #Cas9
- #CRISPR
- Engineering high-fidelity Cas9 often reduces cleavage activity, presenting a challenge for clinical applications.
- Extending the spacer region to 21 or 22 nucleotides restores cleavage activity in SuperFi-Cas9, a high-fidelity variant.
- Cryo-EM structures show that mutations in the RuvC domain create repulsive forces, destabilizing the nuclease-sgRNA-DNA complex.
- Spacer extension enhances PAM-distal interactions, balancing editing efficiency with specificity.
- A deep learning model, AIdit-SuperFi, predicts optimal sgRNA length for high-fidelity genome editing.
- The study provides a strategy to enhance CRISPR complex stability and insights into high-fidelity Cas9's impaired activity.
- Westlake Genetech and Westlake University share intellectual property based on the findings.