Methods in molecular biology (Clifton, N.J.)2027ResearchNon-viral Gene Delivery
1. Rapid Cell-Free Preparation of Circular Vector DNA for pegRNA Expression in Prime Editing
Oliynyk Rt, Church Gm
The expression of prime editing guide RNA (pegRNA) is crucial for effective CRISPR-Cas9 prime genome editing, traditionally facilitated by DNA plasmids, which require a time-consuming process of bacterial cloning, validation, and purification. The Circular Vector (CV) protocol represents an efficient, fully…
- Bacterial cloning
- CRISPR-Cas9
- Circular vector DNA
- DNA ligation
- Gene expression vector
- Prime editing
- Restriction enzyme