（通訊員 阮進學 熊友才）近日，永利動物遺傳育種團隊研究成果以“EXPERT expands prime editing efficiency and range of large fragment edits”為題在Nature Communications發表。研究開發了新型基因編輯工具EXPERT，創新性地在pegRNA nick所在DNA鍊上遊引入額外切口，并對pegRNA末端進行優化改造，同時引入突變以抑制遊離片段對新鍊合成的幹擾，大幅度拓寬了PE編輯範圍，在多個物種中實現了大片段精準高效編輯，為生物育種、人類疾病治療等領域提供了新技術和工具。
▲EXPERT技術原理模式圖
Prime Editing（PE）作為一種精準基因編輯技術，相較于BE（Base Editing）技術具有堿基替換靈活、編輯種類更多、更精準等優點，但也存在編輯範圍受限、長片段編輯效率較低、在豬等動物中編輯效率低等不足，這在一定程度上制約了PE技術的發展和應用。針對這些關鍵問題，永利動物遺傳育種團隊成功研發出EXPERT基因編輯技術，該技術創新性地通過在pegRNA nick所在DNA鍊上遊引入額外切口、對pegRNA末端改造以及引入突變抑制遊離片段對新鍊合成幹擾等手段進行優化，在多個物種中實現了大片段高效精準編輯。
EXPERT相比傳統PE系統具有三個主要優勢：一是更廣泛的編輯範圍，EXPERT能夠同時編輯pegRNA nick的上遊和下遊區域，可實現100bp以上片段高效精準編輯，極大地拓寬了傳統PE編輯範圍。二是更高的編輯效率，與PE2系統相比，EXPERT在長片段（插入、删除、替換）編輯效率最高提升達122倍，同時保持較低的非預期indel比率，實現了大片段高效精準編輯。三是在豬等動物中具有更高的編輯效率，傳統的PE系統在豬中活性不高，EXPERT在豬、鼠、人中都具有很高的編輯效率，顯示出良好跨物種基因編輯能力，為生物育種和人類疾病治療提供了新技術和工具。
yl7703永利動科動醫學院博士畢業生熊友才、博士研究生蘇寅钰和何瑞高為該論文共同第一作者，趙書紅教授、阮進學副研究員和李新雲教授為共同通訊作者。該研究得到了國家重點研發項目、國家自然科學基金創新群體項目等項目資助。
審核人：李新雲
【英文摘要】
Prime editing systems (PEs) hold great promise in modern biotechnology. However, their editing range is limited as PEs can only modify the downstream sequences of the pegRNA nick. Here, we report the development of the extended prime editor system (EXPERT) to overcome this limitation by using an extended pegRNA (ext-pegRNA) with modified 3’ extension, and an additional sgRNA (ups-sgRNA) targeting the upstream region of the ext-pegRNA. We demonstrate that EXPERT can efficiently perform editing on both sides of the ext-pegRNA nick, a task that is unattainable by canonical PEs. EXPERT exhibits prominent capacity in replacing sequences up to 88 base pairs and inserting sequences up to 100 base pairs within the upstream region of the ext-pegRNA nick. Compared to canonical PEs such as PE2, the utilization of the EXPERT strategy significantly enhances the editing efficiency for large fragment edits with an average improvement of 3.12-fold, up to 122.1 times higher. Safety wise, the use of ups-sgRNA does not increase the rates of undesirable insertions and deletions (indels), as the two nicks are on the same strand. Moreover, we do not observe increased off-target editing rates genome-wide. Our work introduces EXPERT as a PE tool with significant potential in life sciences.
論文鍊接：https://www.nature.com/articles/s41467-025-56734-9?utm_source=rct_congratemailt&utm_medium=email&utm_campaign=oa_20250213&utm_content=10.1038/s41467-025-56734-9
