Prime editing-mediated microhomology enables efficient replacement of large DNA - PubMed
5 hours ago
- #genome editing
- #prime editing
- #DNA replacement
- Prime Editing-Microhomology-Enabled Replacement (PREMIER) is a DSB-free platform enabling efficient replacement of large DNA segments.
- PREMIER uses single-stranded microhomology arms at donor and genomic junctions installed via Prime Editing, bypassing the need for long homology arms.
- In cell lines, PREMIER achieved a mean efficiency of 63.4%, exceeding homology-directed repair by 10-20-fold and reducing off-target integrations by over 100-fold compared to nonhomologous end joining.
- The platform simplifies donor preparation and achieves targeted replacement of sequences up to 10.3 kb, including integration of a 6.2-kb oncogene cassette in mouse liver in vivo.
- PREMIER replaced murine Trp53 with human TP53 CDS, generating functional humanized mice, offering a precise tool for complex modeling and therapeutic genome editing.