Journal Article

Enhancement of prime editing via xrRNA motif-joined pegRNA

Liu, Jianghuai, Liu, Yao, Huang, Xingxu, Wang, Xiaolong, Ji, Quanjiang, Qu, Shiyuan, Yao, Yuan, Zhang, Guiquan, Cheng, Daolin, Huang, Shisheng

2022

Overview

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 ( xr RNA) 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.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13 / 13/1 / 13/106 / 13/109 / 13/31 / 13/44 / 38/22 / 38/23 / 38/71 / 38/77 / 38/88 / 38/90 / 42 / 42/41 / 631/1647/1513/1967/3196 / 631/208/4041/3196 / Cell Line / Cell lines / CRISPR-Cas Systems - genetics / Editing / Gene Editing / Genome / Genomes / Humanities and Social Sciences / multidisciplinary / Science / Science (multidisciplinary)