DNA Origami-Platelet Adducts: Nanoconstruct Binding without Platelet Activation.
DNA Origami-Platelet Adducts: Nanoconstruct Binding without Platelet Activation.
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DNA折纸骨骼加合物：纳米结构结合而无需血小板激活。
DOI：
10.1021/acs.bioconjchem.2c00197
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发表时间：
2022-07-20
影响因子：
4.7
通讯作者：
Taylor, Rebecca E.
中科院分区：
文献类型：
--
作者：
Roka-Moiia, Yana;Walawalkar, Vismaya;Liu, Ying;Italiano, Joseph E.;Slepian, Marvin J.;Taylor, Rebecca E.
血小板是一种小型的机械敏感血细胞，负责维持血管完整性，并可根据需要激活以限制出血并促进血栓形成。在血液中循环时，血小板暴露于一系列机械和化学刺激，血小板膜是由外向内信号传递的主要界面和传感器。感测和调节这些界面信号将有助于研究机械化学相互作用；然而，迄今为止，尚未确定将用于传感器制造的加合物附着到血小板上而不触发血小板激活的方法。我们假设 DNA 折纸及其附着方法可以进行优化，以实现血小板膜的非激活仪器。我们设计并制造了多价 DNA 折纸纳米瓦结构，以研究纳米瓦与膜嵌入的单链 DNA-四甘醇胆固醇接头的杂交。开发并验证了两种杂交方案（方法 I 和 II），用于实现 DNA 折纸纳米片与人血小板的高密度结合。使用定量流式细胞术，我们发现当结合突出端的数量从两个增加到六个时，DNA 折纸结合效率显着提高。然而，当将纳米瓦突出的数量进一步增加到 12 时，没有观察到额外的结合益处。使用流式细胞术和透射电子显微镜，我们验证了与 DNA 折纸结构的杂交不会引起血小板形态、活化、聚集或血小板衍生微粒生成的改变。在此，我们证明可以用 DNA 折纸构建体成功地对血小板进行检测，而对血小板形态和功能没有影响或影响很小。我们的方案允许使用少量 DNA 材料将 DNA 折纸与血小板高效高密度结合，从而及时标记大量血小板。非活化血小板-纳米颗粒加合物为推进用于细胞粘附机械传感和治疗剂递送的 DNA 折纸纳米结构的开发提供了一条途径。
Platelets are small, mechanosensitive blood cells responsible for maintaining vascular integrity and activatable on demand to limit bleeding and facilitate thrombosis. While circulating in the blood, platelets are exposed to a range of mechanical and chemical stimuli, with the platelet membrane being the primary interface and transducer of outside-in signaling. Sensing and modulating these interface signals would be useful to study mechanochemical interactions; yet, to date, no methods have been defined to attach adducts for sensor fabrication to platelets without triggering platelet activation. We hypothesized that DNA origami, and methods for its attachment, could be optimized to enable nonactivating instrumentation of the platelet membrane. We designed and fabricated multivalent DNA origami nanotile constructs to investigate nanotile hybridization to membrane-embedded single-stranded DNA-tetraethylene glycol cholesteryl linkers. Two hybridization protocols were developed and validated (Methods I and II) for rendering high-density binding of DNA origami nanotiles to human platelets. Using quantitative flow cytometry, we showed that DNA origami binding efficacy was significantly improved when the number of binding overhangs was increased from two to six. However, no additional binding benefit was observed when increasing the number of nanotile overhangs further to 12. Using flow cytometry and transmission electron microscopy, we verified that hybridization with DNA origami constructs did not cause alterations in the platelet morphology, activation, aggregation, or generation of platelet-derived microparticles. Herein, we demonstrate that platelets can be successfully instrumented with DNA origami constructs with no or minimal effect on the platelet morphology and function. Our protocol allows for efficient high-density binding of DNA origami to platelets using low quantities of the DNA material to label a large number of platelets in a timely manner. Nonactivating platelet–nanotile adducts afford a path for advancing the development of DNA origami nanoconstructs for cell-adherent mechanosensing and therapeutic agent delivery.
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影响因子：
4.6
作者：
Lasne D;Pascreau T;Darame S;Bourrienne MC;Tournoux P;Philippe A;Ziachahabi S;Suarez F;Marcais A;Dupont A;Denis CV;Kauskot A;Borgel D
通讯作者：
Borgel D
影响因子：
37.8
作者：
Kleinschnitz, Christoph;Pozgajova, Miroslava;Stoll, Guido
通讯作者：
Stoll, Guido
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14.9
作者：
Douglas SM;Marblestone AH;Teerapittayanon S;Vazquez A;Church GM;Shih WM
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Shih WM
影响因子：
3.9
作者：
Khmelinskaia, Alena;Muecksch, Jonas;Schwille, Petra
通讯作者：
Schwille, Petra
影响因子：
6.7
作者：
Liu Y ;Wijesekara P ;Kumar S ;Wang W ;Ren X ;Taylor RE
通讯作者：
Taylor RE