CRISPR-associated transposon for programmable viral vector engineering and prime editing - PubMed
9 hours ago
- #Baculovirus
- #Prime Editing
- #CRISPR
- SHOT 2.0 is an optimized CRISPR-associated transposon platform for programmable bacmid engineering.
- It enables RNA-guided and customized bacmid editing in Escherichia coli for producing recombinant baculoviruses.
- Supports site-specific integration of large DNA cargos (up to 14 kb) into defined loci like v-cath and ODVe56.
- Integration at ODVe56 improves transgene stability during serial virus passaging.
- Fully compatible with the Bac-to-Bac® workflow, allowing dual-gene insertion into bacmid and derived baculovirus.
- An all-in-one baculovirus encoding the PE5max prime editor was constructed using SHOT 2.0.
- Achieves prime editing efficiencies up to 85.6% in HEK293T cells and 37.1% in hard-to-transfect cells like iPSCs and liver cancer cells.
- SHOT 2.0 expands the baculovirus engineering toolbox for genome editing and gene delivery.