Wang Dong-Xia, Wang Jing, Wang Ya-Xin, Ma Jia-Yi, Liu Bo, Tang An-Na, Kong De-Ming
State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University Tianjin 300071 P. R. China
School of Medical Laboratory, College of Medical Technology, Tianjin Medical University Guangdong Road Tianjin 300203 P. R. China.
Chem Sci. 2022 Aug 12;13(35):10395-10405. doi: 10.1039/d2sc03374g. eCollection 2022 Sep 14.
The separation and detection of circulating tumor cells (CTCs) have a significant impact on clinical diagnosis and treatment by providing a predictive diagnosis of primary tumors and tumor metastasis. But the responsive release and downstream analysis of live CTCs will provide more valuable information about molecular markers and functional properties. To this end, specific capture and controllable release methods, which can achieve the highly efficient enrichment of CTCs with strong viability, are urgently needed. DNA networks create a flexible, semi-wet three-dimensional (3D) microenvironment for cell culture, and have the potential to minimize the loss of cell viability and molecular integrity. More importantly, responsive DNA networks can be reasonably designed as smart sensors and devices to change shape, color, disassemble, and giving back to external stimuli. Here, a strategy for specifically collecting cells using a dual-aptamer DNA network is designed. The proposed strategy enables effective capture, 3D encapsulation, and responsive release of CTCs with strong viability, which can be used for downstream analysis of live cells. The programmability of CRISPR/Cas12a, a powerful toolbox for genome editing, is used to activate the responsive release of captured CTCs from the DNA network. After activation by a specified double-strand DNA (dsDNA) input, CRISPR/Cas12a cleaves the single-stranded DNA regions in the network, resulting in molecular to macroscopic changes in the network. Accompanied by the deconstruction of the DNA network into fragments, controllable cell release is achieved. The viability of released CTCs is well maintained and downstream cell analysis can be performed. This strategy uses the enzymatic properties of CRISPR/Cas12a to design a platform to improve the programmability and versatility of the DNA network, providing a powerful and effective method for capturing and releasing CTCs from complex physiological samples.
循环肿瘤细胞（CTC）的分离与检测通过对原发性肿瘤和肿瘤转移进行预测性诊断，对临床诊断和治疗具有重大影响。但是，活CTC的响应性释放和下游分析将提供有关分子标记和功能特性的更有价值的信息。为此，迫切需要能够实现具有高活力的CTC高效富集的特异性捕获和可控释放方法。DNA网络为细胞培养创造了一个灵活的半湿三维（3D）微环境，并有可能将细胞活力和分子完整性的损失降至最低。更重要的是，响应性DNA网络可以合理设计为智能传感器和设备，以改变形状、颜色、拆解并响应外部刺激。在此，设计了一种使用双适体DNA网络特异性收集细胞的策略。所提出的策略能够有效捕获、3D封装并响应性释放具有高活力的CTC，可用于活细胞的下游分析。CRISPR/Cas12a作为基因组编辑的强大工具箱，其可编程性被用于激活从DNA网络中捕获的CTC的响应性释放。在特定双链DNA（dsDNA）输入激活后，CRISPR/Cas12a切割网络中的单链DNA区域，导致网络从分子到宏观的变化。伴随着DNA网络解构为片段，实现了可控的细胞释放。释放的CTC的活力得到很好的维持，并且可以进行下游细胞分析。该策略利用CRISPR/Cas12a的酶促特性设计了一个平台，以提高DNA网络的可编程性和通用性，为从复杂生理样本中捕获和释放CTC提供了一种强大而有效的方法。
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