该课题组发现桥接重组酶ISCro4在人类细胞中高度活跃，并提供了其增强活性的结构见解。使用质粒或全RNA为基础的递送，ISCro4支持可编程的千碱基切除和逆转录，并促进供体DNA在基因组位点的插入，效率超过6%。最后，该团队评估了ISCro4的特异性和脱靶活性。这些结果为桥接重组酶作为下一代编辑工具的开发建立了框架，这些工具超出了当前技术的能力。
研究人员表示，基因大小的DNA片段的位点特异性插入在基因组编辑领域仍然是一个未满足的需求。IS110家族丝氨酸重组酶最近被证明在细菌中介导可编程DNA重组，其主题是双特异性RNA向导（桥RNA），同时识别靶点和供体位点。
附：英文原文
Title: Programmable genome editing in human cells using RNA-guided bridge recombinases
Author: Oana Pelea, András Tálas, Javier Fernández Carrera, Nicolas Mathis, Lilly van de Venn, Charles D. Yeh, Péter I. Kulcsár, Kim F. Marquart, Yanik Weber, Saskia E. Gerecke, Isabelle F. Harvey-Seutcheu, Dominic Mailnder, Moritz M. Pfleiderer, Christelle Chanez, Jacob E. Corn, Gerald Schwank, Martin Jinek
Issue&Volume: 2026-02-05
Abstract: Site-specific insertion of gene-sized DNA fragments remains an unmet need in the genome editing field. IS110-family serine recombinases have recently been shown to mediate programmable DNA recombination in bacteria using a bispecific RNA guide (bridge RNA) that simultaneously recognizes target and donor sites. Here, we show that the bridge recombinase ISCro4 is highly active in human cells, and provide structural insights into its enhanced activity. Using plasmid- or all-RNA-based delivery, ISCro4 supports programmable multi-kilobase exisions and inversions, and facilitates donor DNA insertion at genomic sites with efficiencies exceeding 6%. Finally, we assess ISCro4 specificity and off-target activity. These results establish a framework for the development of bridge recombinases as next-generation tools for editing modalities that are beyond the capabilities of current technologies.
DOI: adz1884
Source: https://www.science.org/doi/10.1126/science.adz1884