Multiplexed, precise genome engineering in monocots with twin prime editing systems - PubMed
4 hours ago
- #genome engineering
- #monocots
- #prime editing
- Twin prime editing-based knockout (TKO) system installs stop codon clusters for precise translational termination with minimal in-frame mutations.
- TKO achieves knockout efficiencies up to 70.5% in rice, 58.6% in maize, and 75.1% in wheat protoplasts, and produces heritable knockouts in 96.8% of regenerated rice plants.
- In hexaploid wheat, TKO outperforms Cas9 4.2-fold in generating triple-homolog knockouts by reducing in-frame mutations.
- Orthogonal TKO editors with sequence-divergent SCCs enable simultaneous knockout of up to ten genes without cross-interference.
- Integration of TKO with conventional prime editing forms TRIM1 for simultaneous knockout and precise editing, achieving 22.8% coediting of four genes in rice.
- TRIM2 extends modifications to kilobase-scale via a prime editor-recombinase system, enabling 4.9-kb insertion (1.2% efficiency) and knockout up to 79.8% in protoplasts.