Structural basis of supercoiling-induced CRISPR-Cas9 off-target activity - PubMed
5 hours ago
- #genome-editing
- #structural biology
- #CRISPR-Cas9
- CRISPR-Cas9 is a genome-editing tool with potential off-target activity issues.
- Negative supercoiling ((-)SC) DNA minicircles reveal structural defects resolved by Cas9 binding.
- Cryo-electron microscopy shows Cas9's HNH domain in a more catalytically active conformation in off-target configurations.
- New DNA-RNA mismatch geometries are observed across the protospacer.
- Structural plasticity in the protospacer adjacent motif distal region depends on topology.
- The study provides insights for designing next-generation high-fidelity CRISPR effectors considering topological context.