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首页> 外文期刊>RSC Advances >Proximity hybridization triggered strand displacement and DNAzyme assisted strand recycling for ATP fluorescence detection in vitro and imaging in living cells
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Proximity hybridization triggered strand displacement and DNAzyme assisted strand recycling for ATP fluorescence detection in vitro and imaging in living cells

机译：接近杂交引发的链位移和DNAzyme辅助链回收用于在活细胞体外进行ATP荧光检测的循环回收

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摘要

We developed a novel strategy for ATP detection in vitro and imaging in living cells based on integrating proximity hybridization-induced strand displacement and metal ion-dependent DNAzyme recycling amplification. Four DNA oligonucleotides were used in the sensing system including two aptamer probes, enzymatic sequences and FAM-linked substrate strands. Upon the addition of ATP, the proximity binding of two aptamers to ATP led to the release of the enzymatic sequences, which hybridized with the FAM-linked substrate strand on the graphene oxide (GO) surface to form the ion-dependent DNAzyme. Subsequent catalytic cleavage of the DNAzyme by the corresponding metal ions results in recycling of the enzymatic sequences and cyclic cleavage of the substrate strand, liberating many short FAM-linked oligonuleotide fragments separated from the GO surface, which results in fluorescence enhancement due to the weak affinity of the short FAM-linked oligonuleotide fragment to GO. The amount of produced short FAM-linked oligonuleotide fragments is positively related to the concentration of ATP. This means that one target binding could result in cleaving multiplex fluorophore labelled substrate strands, which provided effective signal amplification. The vivo studies suggested that the nanoprobe was efficiently delivered into living cells and worked for specific, high-contrast imaging of target ATP. More importantly, this target-responsive nanoscissor model is an important approach for intracellular amplified detection and imaging of various analytes by selecting appropriate affinity ligands.
机译：我们在基于整合杂交杂交诱导的链位移和金属离子依赖性DNAzyme回收扩增的活细胞体外和成像中开发了一种新的ATP检测策略。在传感系统中使用四种DNA寡核苷酸，包括两个适体探针，酶序列和FAM连接的衬底股线。在加入ATP，接近到ATP 2点的适体结合导致酶序列的释放，这与石墨烯氧化物（GO）表面上的FAM联基质链杂交以形成所述离子依赖性DNA核酶。随后通过相应的金属离子的DNAzyme催化切割导致酶序列的再循环和衬底股的循环裂解，从转体表面分离出许多短的FAM连接的寡核苷酸片段，这导致由于弱亲和力引起的荧光增强短的Fam-Linked oligonuleotide片段去。产生的短FAM连接的寡核苷酸片段的量与ATP的浓度正相关。这意味着一种目标结合可以导致裂解多重荧光团标记的衬底股线，其提供有效的信号放大。体内研究表明，NanoProbe有效地递送到活细胞中，并为靶ATP的特异性高对比度成像工作。更重要的是，该靶响应纳米镜模型是通过选择合适的亲和配体来细胞内扩增和成像的重要方法。

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  • 来源
    《RSC Advances》 |2018年第49期|共11页
  • 作者

    Gao Fenglei; Wu Jing; Yao Yao; Zhang Yu; Liao Xianjiu; Geng Deqin; Tang Daoquan;

  • 作者单位

    Xuzhou Med Univ Sch Pharm Jiangsu Key Lab New Drug Res &

    Clin Pharm Xuzhou 221004 Jiangsu Peoples R China;

    Xuzhou Med Univ Sch Pharm Jiangsu Key Lab New Drug Res &

    Clin Pharm Xuzhou 221004 Jiangsu Peoples R China;

    Xuzhou Med Univ Sch Pharm Jiangsu Key Lab New Drug Res &

    Clin Pharm Xuzhou 221004 Jiangsu Peoples R China;

    Xuzhou Med Univ Sch Pharm Jiangsu Key Lab New Drug Res &

    Clin Pharm Xuzhou 221004 Jiangsu Peoples R China;

    Youjiang Med Univ Nationalities Sch Pharm Baise 533000 Peoples R China;

    Xuzhou Med Univ Sch Pharm Jiangsu Key Lab New Drug Res &

    Clin Pharm Xuzhou 221004 Jiangsu Peoples R China;

    Xuzhou Med Univ Sch Pharm Jiangsu Key Lab New Drug Res &

    Clin Pharm Xuzhou 221004 Jiangsu Peoples R China;

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  • 正文语种 eng
  • 中图分类 化学;
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