Wrapping DNA-gated mesoporous silica nanoparticles for quantitative monitoring of telomerase activity with glucometer readout.
Wrapping DNA-gated mesoporous silica nanoparticles for quantitative monitoring of telomerase activity with glucometer readout.
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DOI：
10.1039/c4tb00843j
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发表时间：
2014-08
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影响因子：
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通讯作者：
Youmei Wang;Minghua Lu;Jinhua Zhu;Shufang Tian
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文献类型：
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作者：
Youmei Wang;Minghua Lu;Jinhua Zhu;Shufang Tian
本研究报告了一种简单而灵敏的端粒酶活性定量监测方案，该方案基于包裹dna的介孔二氧化硅纳米颗粒（MSNs）的靶反应释放，通过与便携式个人血糖仪（PGM）耦合。为了构建这样的分析系统，葡萄糖分子最初被加载到胺化的MSN的孔中，然后用特殊设计的包裹DNA密封孔。添加端粒酶和dNTPs后，在端粒酶的帮助下，组装的包裹DNA链在胺化的MSN上被延长。随着端粒酶的进展，延伸的DNA链从MSN分离，由于形成刚性的，发夹状的DNA结构。然后“分子门”被打开，导致葡萄糖从MSN中释放出来。释放的葡萄糖分子可以使用外部PGM进行定量监测。PGM信号随着端粒酶活性的增加而增加。在最佳条件下，基于pgm的传感平台显示出良好的端粒酶活性分析特性，并且可以在低至80个细胞mL-1的浓度下检测HeLa提取物中的端粒酶活性。利用体细胞和肿瘤细胞系，对该试验的一般性进行了评估，结果令人满意。3’-叠氮-3’-脱氧胸腺嘧啶的抑制作用在端粒酶抑制剂筛选研究中也得到了很好的表现。该方法重复性好，操作简便，为临床端粒酶活性定量测定提供了一种实用的方法。
This work reports a simple and sensitive sensing protocol for the quantitative monitoring of telomerase activity based on target-responsive release of cargo from wrapping DNA-capped mesoporous silica nanoparticles (MSNs) by coupling with a portable personal glucometer (PGM). To construct such an assay system, glucose molecules are initially loaded into the pores of the aminated MSN, and the pores are then sealed with a specially designed wrapping DNA. Upon the addition of telomerase and dNTPs, the assembled wrapping DNA strands are prolonged with the assistance of telomerase on the aminated MSN. Accompanying the progression of telomerase, the extended DNA strands detach from the MSN, owing to the formation of rigid, hairpin-like DNA structures. The "molecular gates" are then opened, resulting in the release of glucose from MSN. The released glucose molecules can be quantitatively monitored using an external PGM. The PGM signal increases with the increment of telomerase activity. Under optimal conditions, the PGM-based sensing platform exhibits good analytical properties for the determination of telomerase activity and allows for the detection of telomerase activity in the HeLa extract at concentrations as low as 80 cells mL-1. Using somatic and tumor cell lines, the generality of the assay is evaluated with satisfactory results. The inhibition effect of 3'-azido-3'-deoxythymidine also receives a good performance in the telomerase-inhibitor screening research. The methodology affords good reproducibility and simple operations, thus providing a useful scheme for practical use in a quantitative telomerase activity assay for clinical application.