Determining the Cytosolic Stability of Small DNA Nanostructures In Cellula.
Determining the Cytosolic Stability of Small DNA Nanostructures In Cellula.
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确定细胞内小型DNA纳米结构的胞质稳定性
DOI：
10.1021/acs.nanolett.2c00917
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发表时间：
2022-06-22
期刊：
影响因子：
10.8
通讯作者：
Medintz, Igor L.
中科院分区：
文献类型：
--
作者：
Mathur, Divita;Rogers, Katherine E.;Diaz, Sebastian A.;Muroski, Megan E.;Klein, William P.;Nag, Okhil K.;Lee, Kwahun;Field, Lauren D.;Delehanty, James B.;Medintz, Igor L.
DNA纳米结构已被证明在生物医学方面具有潜力。然而，它们在细胞内的相互作用——尤其是胞质稳定性——仍然是未知的，并且由于内吞摄取和捕获的复杂性，试图辨别这一点的努力受到了干扰。在这里，我们绕过内吞摄取，直接评估活细胞中的DNA结构稳定性。用多步Förster共振能量转移染料级联标记常用的DNA结构-十字和四面体，并将其微注射到转化细胞和原代细胞的细胞质中。荧光显微镜监测的能量转移损失报告了该结构在细胞内的直接时间分辨分解。结果显示，DNA十字准星在20分钟内迅速降解，而四面体在注射后至少1小时内保持完整。核酸酶分析结合当前对四面体扭转刚度的理解证实了其较高的稳定性。这些研究可以为未来需要可编程降解率的DNA纳米结构的设计参数提供信息。
DNA nanostructures have proven potential in biomedicine. However, their intracellular interactions—especially cytosolic stability—remain mostly unknown and attempts to discern this are confounded by the complexities of endocytic uptake and entrapment. Here, we bypass the endocytic uptake and evaluate the DNA structural stability directly in live cells. Commonly used DNA structures—crosshairs and a tetrahedron—were labeled with a multistep Förster resonance energy transfer dye cascade and microinjected into the cytosol of transformed and primary cells. Energy transfer loss, as monitored by fluorescence microscopy, reported the structure’s direct time-resolved breakdown in cellula. The results showed rapid degradation of the DNA crosshair within 20 min, while the tetrahedron remained consistently intact for at least 1 h postinjection. Nuclease assays in conjunction with a current understanding of the tetrahedron’s torsional rigidity confirmed its higher stability. Such studies can inform design parameters for future DNA nanostructures where programmable degradation rates may be required.
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影响因子：
17.1
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影响因子：
4.7
作者：
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通讯作者：
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影响因子：
48
作者：
Algar, W. Russ;Hildebrandt, Niko;Medintz, Igor L.
通讯作者：
Medintz, Igor L.