CRISPR-Cas9已经被探索作为下调靶基因的治疗药物，控制递送Cas9核糖核蛋白（RNP）对治疗效果至关重要，但仍然存在挑战。在这里，我们报道了DNA纳米结构（NF）的级联动态装配/解聚，实现了对Cas9 RNP的控制递送。NF是用丙烯酰胺修饰的DNA制备而成，启动了级联杂交链反应（HCR）。通过HCR，将单导RNA并入到NF中，同时扩张了NF的内部空间，方便Cas9蛋白的装载。NF设计了亲水性酰胺和疏水性异丙基，实现了动态肿胀和聚集。通过引入含有二硫键的N,N-双（丙烯酰基）胞嘧胺， NF能够响应癌细胞内的谷胱甘肽引发完全解聚。体外和体内研究结果表明，该方法在癌细胞中具有高基因编辑效率，在正常细胞中具有低毒性，在乳腺癌小鼠模型中具有明显的抗肿瘤效果。

CRISPR-Cas9 has been explored as a therapeutic agent for down-regulating target genes; the controlled delivery of Cas9 ribonucleoprotein (RNP) is essential for therapeutic efficacy and remains a challenge. Here, we report cascade dynamic assembly/disassembly of DNA nanoframework (NF) that enables the controlled delivery of Cas9 RNP. NF was prepared with acrylamide-modified DNA that initiated cascade hybridization chain reaction (HCR). Through an HCR, single-guide RNA was incorporated to NF; simultaneously, the internal space of NF was expanded, facilitating the loading of Cas9 protein. NF was designed with hydrophilic acylamino and hydrophobic isopropyl, allowing dynamic swelling and aggregation. The responsive release of Cas9 RNP was realized by introducing disulfide bond-containing N,N-bis(acryloyl)cystamine that was specifically in response to glutathione of cancer cells, triggering the complete disassembly of NF. In vitro and in vivo investigations demonstrated the high gene editing efficiency in cancer cells, the hypotoxicity in normal cells, and notable antitumor efficacy in a breast cancer mouse model.