Increasing the efficiency and precision of prime editing with guide RNA pairs.

Abstract

The recently reported prime editor (PE) can produce all types of base substitution, insertion and deletion, greatly expanding the scope of genome editing. However, improving the editing efficiency and precision of PE represents a major challenge. Here, we report an approach termed the homologous 3' extension mediated prime editor (HOPE). HOPE uses paired prime editing guide RNAs (pegRNAs) encoding the same edits in both sense and antisense DNA strands to achieve high editing efficiency in human embryonic kidney 293T cells as well as mismatch repair-deficient human colorectal carcinoma 116 cells. In addition, we found that HOPE shows greatly improved product purity compared to the original PE3 system. We envision that this enhanced tool could broaden both fundamental research and therapeutic applications of prime editing.

• Authors: Yuan Zhuang, Jiangle Liu, Hao Wu, Qingguo Zhu, Yongchang Yan, Haowei Meng, Peng R Chen, Chengqi Yi
• Journal: Nature chemical biology (Nat Chem Biol) Vol. 18 Issue 1 Pg. 29-37 (01 2022) ISSN: 1552-4469 [Electronic] United States
• PMID: 34711981 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.

Chemical References

• RNA, Guide

Topics

• CRISPR-Cas Systems
• HEK293 Cells
• Humans
• RNA Editing
• RNA, Guide, Kinetoplastida (genetics)