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EXPERT expands prime editing efficiency and range of large fragment edits
Youcai Xiong1,2, Yinyu Su1,2, Ruigao He1,2
1Frontiers Science Center for Animal Breeding and Sustainable Production, Huazhong Agricultural University, Wuhan, PR China.
Nature Communications
|February 12, 2025
Summary
Extended prime editing systems (EXPERT) expand gene editing capabilities beyond current limitations. This novel approach enables precise DNA modifications on both sides of a nick, significantly improving efficiency for large edits without increasing off-target effects.
Area of Science:
- Molecular Biology
- Gene Editing Technologies
- Biotechnology
Background:
- Prime editing systems (PEs) offer precise genome editing but are limited by their editing range.
- Canonical PEs can only modify sequences downstream of the nick site introduced by the pegRNA.
Purpose of the Study:
- To develop an extended prime editor system (EXPERT) to overcome the range limitations of canonical PEs.
- To enable efficient editing on both sides of the nick site.
Main Methods:
- Development of an extended prime editor system (EXPERT) utilizing an extended pegRNA (ext-pegRNA) and an additional upstream-targeting sgRNA (ups-sgRNA).
- Demonstration of EXPERT's ability to perform edits on both upstream and downstream regions of the nick.
Main Results:
- EXPERT efficiently replaces sequences up to 88 bp and inserts sequences up to 100 bp in the upstream region.
- EXPERT significantly enhances editing efficiency for large fragment edits (average 3.12-fold improvement, up to 122.1 times higher than PE2).
- EXPERT does not increase indel rates or genome-wide off-target editing events.
Conclusions:
- EXPERT expands the editing capacity of prime editing systems.
- This novel system offers a powerful tool for precise and efficient large-scale genome editing in life sciences.
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