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Three-Dimensional DNA Nanomachine Biosensor by Integrating DNA Walker and Rolling Machine Cascade Amplification for Ultrasensitive Detection of Cancer-Related Gene.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-07-09 , DOI: 10.1021/acs.analchem.0c01074 Na Wu 1 , Kun Wang 1 , Yi-Ting Wang 1 , Ming-Li Chen 1 , Xu-Wei Chen 1 , Ting Yang 1 , Jian-Hua Wang 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-07-09 , DOI: 10.1021/acs.analchem.0c01074 Na Wu 1 , Kun Wang 1 , Yi-Ting Wang 1 , Ming-Li Chen 1 , Xu-Wei Chen 1 , Ting Yang 1 , Jian-Hua Wang 1
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Stochastic DNA walkers capable of traversing on three-dimensional (3D) tracks have received great deal of attention. However, DNA walker-based biosensors exhibit limited amplification efficiency because of their slow walking kinetics and low processivity. Herein, by taking advantage of the high processivity of a DNA rolling machine, a sensitive ratiometric DNA nanomachine biosensor is designed. The biosensor is constructed with hairpin-loaded Au nanoparticles (NPs) (hpDNA@AuNPs) as a DNA walker and AgNCs-decorated magnetic NPs (AgNCs@MNPs) as a DNA rolling machine. In the presence of target DNA, exonuclease III (Exo III)-powered DNA walker is activated to accomplish first-stage amplification via a burnt-bridge mechanism, generating a great deal of toehold-loaded AuNPs (Toehold@AuNPs) to hybridize with magnetic nanoparticles loaded with silver-nanoclusters-labeled DNA (AgNCs@MNPs) with the assistance of Exo III. These trigger rapid rolling of AuNPs on the AgNCs@MNPs surface and release free AgNCs, converting the biological signal into a mass spectrometric signal ratio (107Ag/197Au) with detection by ICP-MS. A linear range of 0.5–500 fmol L–1 is achieved with a detection limit of 119 amol L–1 for the p53 gene. The practical applicability of the biosensor has been demonstrated in the accurate assay of the p53 gene in the human blood.
中文翻译：
集成了DNA Walker和Rolling Machine级联扩增的三维DNA纳米机生物传感器，用于超灵敏检测癌症相关基因。
能够在三维（3D）轨道上移动的随机DNA步行者受到了广泛的关注。但是，基于DNA Walker的生物传感器由于其缓慢的行走动力学和低的合成能力而显示出有限的扩增效率。在此，通过利用DNA轧制机的高生产率，设计了灵敏的比例式DNA纳米机生物传感器。该生物传感器由装有发夹的金纳米颗粒（NPs）（hpDNA @ AuNPs）作为DNA Walker和装饰有AgNCs的磁性NP（AgNCs @ MNPs）构成。在目标DNA存在的情况下，由核酸外切酶III（Exo III）驱动的DNA Walker被激活，以通过烧桥机制完成第一步扩增，在Exo III的协助下，产生了大量的脚趾负载AuNP（Toehold @ AuNPs）与载有银纳米簇标记的DNA（AgNCs @ MNPs）的磁性纳米颗粒杂交。这些触发AuNPs在AgNCs @ MNPs表面上快速滚动并释放出游离的AgNCs，从而将生物信号转换为质谱信号比（107 Ag / 197 Au），并通过ICP-MS检测。的0.5-500飞摩尔L A线性范围-1与119 AMOL升的检测限实现-1为p53的基因。生物传感器的实际适用性已在人血中p53基因的准确测定中得到了证明。
更新日期：2020-07-09
中文翻译：
集成了DNA Walker和Rolling Machine级联扩增的三维DNA纳米机生物传感器，用于超灵敏检测癌症相关基因。
能够在三维（3D）轨道上移动的随机DNA步行者受到了广泛的关注。但是，基于DNA Walker的生物传感器由于其缓慢的行走动力学和低的合成能力而显示出有限的扩增效率。在此，通过利用DNA轧制机的高生产率，设计了灵敏的比例式DNA纳米机生物传感器。该生物传感器由装有发夹的金纳米颗粒（NPs）（hpDNA @ AuNPs）作为DNA Walker和装饰有AgNCs的磁性NP（AgNCs @ MNPs）构成。在目标DNA存在的情况下，由核酸外切酶III（Exo III）驱动的DNA Walker被激活，以通过烧桥机制完成第一步扩增，在Exo III的协助下，产生了大量的脚趾负载AuNP（Toehold @ AuNPs）与载有银纳米簇标记的DNA（AgNCs @ MNPs）的磁性纳米颗粒杂交。这些触发AuNPs在AgNCs @ MNPs表面上快速滚动并释放出游离的AgNCs，从而将生物信号转换为质谱信号比（107 Ag / 197 Au），并通过ICP-MS检测。的0.5-500飞摩尔L A线性范围-1与119 AMOL升的检测限实现-1为p53的基因。生物传感器的实际适用性已在人血中p53基因的准确测定中得到了证明。
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