Enhancement of prime editing via xrRNA motif-joined pegRNA

The prime editors (PEs) have shown great promise for precise genome modification. However, their suboptimal efficiencies present a significant technical challenge. Here, by appending a viral exoribonuclease-resistant RNA motif (xrRNA) to the 3 '-extended portion of pegRNAs for their increased resistance against degradation, we develop an upgraded PE platform (xrPE) with substantially enhanced editing efficiencies in multiple cell lines. A pan-target average enhancement of up to 3.1-, 4.5- and 2.5-fold in given cell types is observed for base conversions, small deletions, and small insertions, respectively. Additionally, xrPE exhibits comparable edit:indel ratios and similarly minimal off-target editing as the canonical PE3. Of note, parallel comparison of xrPE to the most recently developed epegRNA-based PE system shows their largely equivalent editing performances. Our study establishes a highly adaptable platform of improved PE that shall have broad implications. The prime editors (PEs) have shown great promise for precise genome modification. Here the authors place a stabilizing viral xrRNA motif to the 3 ' of pegRNAs to enhance editing efficiencies.

Automated summary

This study developed an upgraded prime editor platform (xrPE) by adding a viral xrRNA motif to pegRNAs, resulting in significantly enhanced editing efficiencies in multiple cell lines. The xrPE exhibited similar edit:indel ratios and minimal off-target editing compared to the canonical PE3, and showed comparable editing performance to the epegRNA-based PE system.