靶向介导的 DNA 网络自组装，用于活细胞中 APE1 的灵敏检测和细胞内成像,Chemical Science

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靶向介导的 DNA 网络自组装，用于活细胞中 APE1 的灵敏检测和细胞内成像
Chemical Science ( IF 7.6 ) Pub Date : 2023-02-10 , DOI: 10.1039/d2sc06968g
Xue-Mei Zhou ₁ , Ying Zhuo ₁ , Ruo Yuan ₁ , Ya-Qin Chai ₁

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在此，巨型 DNA 网络由两种功能化的四面体 DNA 纳米结构 (f-TDN) 组装而成，用于脱嘌呤/脱嘧啶核酸内切酶 1 (APE1) 的灵敏检测和细胞内成像以及肿瘤细胞中的基因治疗。令人印象深刻的是，由于发夹的局部浓度高、空间限制效应和巨型 DNA 网络的产生，f-TDN 上的催化发夹组装 (CHA) 反应的反应速率比传统的游离 CHA 反应快得多，这显着增强荧光信号，实现 APE1 的灵敏检测，限度为 3.34 × 10 ^-8 U μL ^-1. 更重要的是，f-TDNs上组装的适体Sgc8可以增强DNA结构对肿瘤细胞的靶向活性，使其无需任何转染试剂即可内吞入细胞，从而实现活细胞内APE1的选择性成像。同时，f-TDN1携带的siRNA在内源性靶点APE1存在下可准确释放促进肿瘤细胞凋亡，实现肿瘤的有效精准治疗。得益于高特异性和敏感性，所开发的DNA纳米结构为癌症的精确诊断和治疗提供了一个极好的纳米平台。

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Target-mediated self-assembly of DNA networks for sensitive detection and intracellular imaging of APE1 in living cells

Herein, giant DNA networks were assembled from two kinds of functionalized tetrahedral DNA nanostructures (f-TDNs) for sensitive detection and intracellular imaging of apurinic/apyrimidinic endonuclease 1 (APE1) as well as gene therapy in tumor cells. Impressively, the reaction rate of the catalytic hairpin assembly (CHA) reaction on f-TDNs was much faster than that of the conventional free CHA reaction owing to the high local concentration of hairpins, spatial confinement effect and production of giant DNA networks, which significantly enhanced the fluorescence signal to achieve sensitive detection of APE1 with a limit of 3.34 × 10⁻⁸ U μL⁻¹. More importantly, the aptamer Sgc8 assembled on f-TDNs could enhance the targeting activity of the DNA structure to tumor cells, allowing it to endocytose into cells without any transfection reagents, which could achieve selective imaging of intracellular APE1 in living cells. Meanwhile, the siRNA carried by f-TDN1 could be accurately released to promote tumor cell apoptosis in the presence of endogenous target APE1, realizing effective and precise tumor therapy. Benefiting from the high specificity and sensitivity, the developed DNA nanostructures provide an excellent nanoplatform for precise cancer diagnosis and therapy.

更新日期：2023-02-11

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