Reducing the inherent auto-inhibitory interaction within the pegRNA enhances prime editing efficiency.

ABSTRACT: Prime editing systems have enabled the incorporation of precise edits within a genome without introducing double strand breaks. Previous studies defined an optimal primer binding site (PBS) length for the pegRNA of ∼13 nucleotides depending on the sequence composition. However, optimal PBS length characterization has been based on prime editing outcomes using plasmid or lentiviral expression systems. In this study, we demonstrate that for prime editor (PE) ribonucleoprotein complexes, the auto-inhibitory interaction between the PBS and the spacer sequence affects pegRNA binding efficiency and target recognition. Destabilizing this auto-inhibitory interaction by reducing the complementarity between the PBS-spacer region enhances prime editing efficiency in multiple prime editing formats. In

SUBMITTER: Ponnienselvan K 

PROVIDER: S-EPMC10359601 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature