(pro)-OpenCRISPR-1-SV40pA + U6-(gRNA)-CAG-EGFP-KASH-bGHpA
OpenCRISPR-1 counterpart to the SpCas9 single-gRNA CAG-EGFP-KASH design. Nuclease AAV expresses AI-designed OpenCRISPR-1 (Profluent Bio) under a custom promoter with SV40pA; companion AAV delivers one U6-driven gRNA and CAG-EGFP-KASH for FANS-compatible nuclear envelope labeling. Canonical SpCas9 sgRNAs are fully compatible — the companion AAV is architecturally identical to its SpCas9 counterpart.
Characteristics
OpenCRISPR-1 is an AI-generated nuclease with 403 mutations from SpCas9, comparable on-target activity (56.4% vs 47.1% median indel), 95% fewer off-target edits, and absence of immunodominant SpCas9 T cell epitopes. Strict NGG PAM requirement. Blunt-end DSBs. CAG-EGFP-KASH provides ubiquitous perinuclear fluorescence for FANS-based nuclei isolation independent of cell type.
Parts
Custom Inserts
Related Designs
Literature Sources
- Ruffolo et al. (2025). Design of highly functional genome editors by modelling CRISPR–Cas sequences. Nature- Ruffolo 2025 OpenCRISPR
- Kumar et al. (2018). The Development of an AAV-Based CRISPR SaCas9 Genome Editing System That Can Be Delivered to Neurons in vivo and Regulated via Doxycycline and Cre-Recombinase. Front Mol Neurosci- Kumar 2018 In Vivo CRISPR
- Platt et al. (2014). CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell- Platt 2014 Cas9 Mice
Available Products
No products currently available with this design. Contact us for custom orders.
Additional Information
Validated in: HEK293 cells (OpenCRISPR-1 characterization); companion vector architecture validated in mouse neurons in vivo
Applications: Single-target CRISPR editing where reduced immunogenicity or off-target profile is preferred over wild-type SpCas9. Drop-in replacement for SpCas9 dual-AAV designs using existing SpCas9-compatible gRNAs. EGFP-KASH enables FANS-based nuclei sorting for downstream single-nucleus omics.