A Single-Enzyme Activated CRISPR-Cas12a Nano System via Subtly Balanced dsDNA for Kinetic-Gated UDG Detection and Spatiotemporal Cellular Imaging - PubMed
4 days ago
- #UDG detection
- #cellular imaging
- #CRISPR-Cas12a
- A Single-Enzyme Activated CRISPR-Cas12a Nano System is developed for UDG detection and cellular imaging.
- The system uses a subtly balanced double-stranded DNA (dsDNA) substrate to gate Cas12a activity until UDG-mediated uracil excision occurs.
- This approach directly converts uracil excision into an amplified CRISPR response without additional enzymatic steps.
- The system achieves a 1840-fold discrimination ratio and a detection limit of 5 × 10^-7 U/mL.
- A genetically encoded variant enables nuclear localization for in situ imaging of endogenous UDG.
- The platform visualizes UDG dynamics across cell cycle phases, enabling spatiotemporal mapping in living cells.
- The work introduces a new activation paradigm for CRISPR-Cas12a via balanced dsDNA for precise molecular sensing.