免疫逃逸DNA供体和重组酶赋能千碱基级写入，这一成果由马萨诸塞州总医院Benjamin P. Kleinstiver团队经过不懈努力而取得。这一研究成果于2026年3月11日发表在国际顶尖学术期刊《自然》上。

通过利用整合的原核病毒主题和可移动遗传元件的机制，课题组人员证明重组酶与免疫回避的环状单链DNA分子兼容，这些分子最好带有重建重组酶识别序列的部分双链区。这种方法，课题组人员将其称为通过核酸合成的大长度模板添加（INSTALL）进行整合的方法，与多种蛋白质和RNA引导的重组酶兼容，用于高保真的千碱基级人类基因组编写。与脂质纳米颗粒递送的dsDNA分子相比，INSTALL通过显著提高耐受性和扩大给药范围，提高重组酶介导的整合效率，支持体内系统性非病毒DNA递送，从而将原代人细胞和小鼠的先天免疫反应降至最低。INSTALL通过将免疫隐身核酸与重组酶协同作用，在没有病毒载体的情况下实现千碱基级整合策略，从而克服了DNA传递和整合方法的基本挑战。

据介绍，利用重组酶将千碱基级DNA序列插入哺乳动物基因组的基因组编辑技术通常需要大双链DNA（dsDNA）供体。然而，dsDNA分子引起有问题和有毒的先天免疫反应，限制了整合效率，并且通常限制了对体外或免疫缺陷环境的适用性。

附：英文原文

Title: Immune evasive DNA donors and recombinases license kilobase-scale writing

Author: Tou, Connor J., Xie, Keqiang, Ferreira da Silva, Joana, Kalailingam, Pazhanichamy, Amar-Lewis, Eliz, Rufino-Ramos, David, Sawyer, William, Eller, Madeline L., Starzyk, Jakob, Majumdar, Ishita, Wang, Jiao, Lee, Danna, Yang, Shaobo, Meis, Ronald J., Dahl, Gary A., Li, Jiahe, Shan, Richard, Artzi, Natalie, Musolino, Patricia L., Wu, Hao, Kleinstiver, Benjamin P.

Issue&Volume: 2026-03-11

Abstract: Genome-editing technologies that use recombinases to insert kilobase-scale DNA sequences into mammalian genomes canonically require large double-stranded DNA (dsDNA) donors1,2. However, dsDNA molecules evoke problematic and toxic innate immune responses, limiting integration efficiencies and generally constraining applicability to ex vivo or immune-deficient contexts. By harnessing mechanisms of integrative prokaryotic viruses and mobile genetic elements, here we demonstrate that recombinases are compatible with immune evasive circular single-stranded DNA molecules optimally bearing a partial-duplex region that reconstitutes the recombinase recognition sequence. This approach, which we term integration through nucleus-synthesized template addition of large lengths (INSTALL), is compatible with diverse protein and RNA-guided recombinases for high-fidelity kilobase-scale human genome writing. INSTALL minimizes innate immune responses in primary human cells and in mice, improving recombinase-mediated integration efficiencies and supporting systemic in vivo non-viral DNA delivery by substantially increasing tolerability and broadening the dosing range compared with lipid nanoparticle-delivered dsDNA molecules. Together, INSTALL overcomes fundamental challenges for DNA delivery and integration methods by synergizing immune-stealth nucleic acids with recombinases to enable kilobase-scale integration strategies without viral vectors.

DOI: 10.1038/s41586-026-10241-z

Source: https://www.nature.com/articles/s41586-026-10241-z