麻省大学陈医学院Wen Xue小组宣布他们的研究显示，与线性DNA供体的初始组装使大基因组插入成为可能。该项研究成果发表在2026年4月29日出版的《自然》上。

在这里，研究小组描述了一种用于插入大DNA供体片段的引物组装（PA）方法，其末端被设计成与双引物编辑（twinPE）产生的皮瓣重叠。该团队的主题PA插入一个或多个重叠的DNA片段，总插入大小从0.1KB ~ 11.1 KB。非同源末端连接抑制剂提高了插入的效率和精度。PA依赖于易于产生的DNA模板，不需要外源DNA依赖的DNA聚合酶的共同递送，并在非循环细胞中进行，表明它独立于典型的同源性定向修复途径。他们的研究表明，PA可以在细胞中启动吉布森样组装，在没有双链DNA断裂、重组酶或同源定向修复的情况下产生基因插入。

据悉，靶向插入大片段DNA在基因组工程和基因治疗中具有广阔的应用前景。但当插入片段超过400个碱基对时，其效率仍然较低。

附：英文原文

Title: Prime assembly with linear DNA donors enables large genomic insertions

Author: Liu, Bin, Petti, Andrew, Zhou, Xuntao, Cheng, Haoyang, Gao, Jenny, Yee, Matthew, Qiao, Youwei, Zhang, Yanjun, Zhou, Lin, Wolfe, Scot A., Jiang, Tingting, Sontheimer, Erik J., Xue, Wen

Issue&Volume: 2026-04-29

Abstract: Targeted insertion of large DNA fragments has promising applications for genome engineering and gene therapy1,2. Twin prime-editing guide RNAs have enabled relatively large insertions, but the efficiency remains low for insertions greater than 400base pairs3,4,5,6. Here we describe a prime assembly (PA) approach for the insertion of large DNA donor fragments, of which the ends are designed to overlap with the flaps generated by twin prime editing (twinPE). We used PA to insert one or multiple overlapping DNA fragments, with total insertion sizes ranging from 0.1kb to 11kb. An inhibitor of non-homologous end joining enhanced both the efficiency and precision of insertions. PA relies on DNA templates that are easily produced, does not require co-delivery of exogenous DNA-dependent DNA polymerases and proceeds in non-cycling cells, suggesting independence from canonical homology-directed repair pathways. Our study demonstrates that PA can initiate Gibson-like assembly in cells to generate gene insertions without double-stranded DNA breaks, recombinases or homology-directed repair.

DOI: 10.1038/s41586-026-10460-4

Source: https://www.nature.com/articles/s41586-026-10460-4