Boosting prime editing with engineered non-canonical pegRNAs - PubMed
5 hours ago
- #Genetic Engineering
- #CRISPR-Cas9
- #Prime Editing
- Prime editing (PE) uses canonical pegRNAs for precise genetic modifications but has weak editing efficiency, limiting therapeutic potential.
- Engineered non-canonical pegRNAs (npegRNAs) integrate reverse transcription template and primer binding site within guide RNA loops to boost PE efficiency.
- npegRNAs enhance precise editing across genomic sites and cell types, improve gene correction in a mouse model, and resist exonuclease degradation.
- PE ribonucleoprotein delivery with npegRNAs achieves up to 26.8-fold higher editing yields than canonical pegRNAs and 5.9-fold over engineered pegRNAs.
- npegRNA-mediated RNPs increase efficiency for installing disease-relevant mutations up to 123-fold in human cell lines, including Jurkat T cells and iPS cells.