DNA纳米结构(DN)已经发现在生物传感中的应用越来越多,药物输送,和治疗学,因为它们的可定制装配,尺寸和形状控制,和简单的功能化。然而,它们有限的细胞摄取和核传递阻碍了它们在这些应用中的有效性。这里,我们证明了应用细胞表面结合作为一种通用策略的潜力,能够快速增强细胞内和核内的DNA递送.通过使用点击化学通过胆固醇锚或细胞表面糖萼靶向质膜,我们观察到包括纳米球在内的三种不同类型的DNs的细胞摄取显着增加2到8倍,纳米棒,和纳米瓷砖,在半小时的短时间内。发现几个因素在调节DNs的摄取中起关键作用,包括它们的几何形状,价,结合部分的定位和间隔。简而言之,纳米球普遍优选用于细胞表面附着和内化。然而,边缘装饰的纳米瓷砖弥补了它们的几何缺陷,并在这两个类别中胜过纳米球。此外,我们通过将它们与细胞培养基和细胞裂解物一起孵育来确认DNs的短期结构稳定性。Further,我们研究了细胞表面结合的DNs的内吞途径,并揭示了它是一个涉及多个途径的相互依存的过程,类似于未修改的DN。最后,我们证明了细胞表面附着的DN在核内递送中表现出显著的增强。我们的发现提出了一种应用,利用细胞表面结合来潜在地克服低细胞摄取的局限性。这可能会加强和扩大工具箱的DNA纳米结构系统的有效细胞和核输送。
DNA nanostructures (DNs) have found increasing use in biosensing, drug delivery, and therapeutics because of their customizable assembly, size and shape control, and facile functionalization. However, their limited cellular uptake and nuclear delivery have hindered their effectiveness in these applications. Here, we demonstrate the potential of applying cell-surface binding as a general strategy to enable rapid enhancement of intracellular and intranuclear delivery of DNs. By targeting the plasma membrane via cholesterol anchors or the cell-surface glycocalyx using click chemistry, we observe a significant 2 to 8-fold increase in the cellular uptake of three distinct types of DNs that include nanospheres, nanorods, and nanotiles, within a short time frame of half an hour. Several factors are found to play a critical role in modulating the uptake of DNs, including their geometries, the valency, positioning and spacing of binding moieties. Briefly, nanospheres are universally preferable for cell surface attachment and internalization. However, edge-decorated nanotiles compensate for their geometry deficiency and outperform nanospheres in both categories. In addition, we confirm the short-term structural stability of DNs by incubating them with cell medium and cell lysate. Further, we investigate the endocytic pathway of cell-surface bound DNs and reveal that it is an interdependent process involving multiple pathways, similar to those of unmodified DNs. Finally, we demonstrate that cell-surface attached DNs exhibit a substantial enhancement in the intranuclear delivery. Our findings present an application that leverages cell-surface binding to potentially overcome the limitations of low cellular uptake, which may strengthen and expand the toolbox for effective cellular and nuclear delivery of DNA nanostructure systems.