Mismatch-Assisted Toehold Exchange Cascades for Magnetic Nanoparticle-based Nucleic Acid Diagnostics.

IF 8.7 Q1 CHEMISTRY, MULTIDISCIPLINARY
JACS Au Pub Date : 2025-09-04 eCollection Date: 2025-09-22 DOI:10.1021/jacsau.5c00985
Rebecca Sack, Joshua Evans, Florian Wolgast, Meinhard Schilling, Thilo Viereck, Petr Šulc, Aidin Lak
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引用次数: 0

Abstract

Sensitive, simple, and rapid detection of nucleic acid sequences at point-of-care settings is still an unmet quest. Magnetic readout assays combined with toehold-mediated strand displacement-based circuits are amplification- and wash-free, essential features for contributing to this demand. Nevertheless, nonenzymatic strand displacement circuits are slow, with low sensitivity for early disease diagnostics. Here, we propose novel mismatch-assisted toehold exchange (MATE) magnetic cascades, wherein magnetic susceptibility increases by dissociation of magnetic nanoparticles (MNPs) from engineered magnetic clusters upon detection of a nucleic acid target in solution. The MATE relies on the generation of an allosteric toehold by spontaneous dissociation to efficiently recycle the target, amplify magnetic signal output, and enhance the assay's kinetics. We show that introducing a mismatch in the allosteric toehold domain enhances the overall declustering kinetics 7-fold, as also confirmed with oxDNA simulations, with the largest effect gained for the mismatch being closest to where the branch migration by the target ends. By integrating MATE into magnetic diagnostics cascades, we demonstrate similar sensitivity in a 12-fold shorter assay time compared to our previous circuit design. Our work makes a major leap toward bringing MNP-based diagnostics much closer to the clinical point-of-care settings by offering a simple, rapid, isothermal, and nonenzymatic assay workflow.

失配辅助脚交换级联磁纳米粒子为基础的核酸诊断。
敏感,简单,快速检测核酸序列在护理点设置仍然是一个未满足的要求。磁读出分析结合了基于支点介导的链位移电路是扩增和免清洗的,是满足这一需求的基本特征。然而,非酶链位移回路是缓慢的,对早期疾病诊断的敏感性较低。在这里,我们提出了一种新的错配辅助支点交换(MATE)磁级联,其中磁性纳米颗粒(MNPs)在检测到溶液中的核酸靶标时从工程磁团簇中解离,从而增加磁化率。MATE依赖于自发解离产生的变弹性支点来有效地回收目标,放大磁信号输出,并增强分析的动力学。我们发现,在变构脚域引入错配可以提高整体脱簇动力学7倍,正如oxDNA模拟所证实的那样,在最接近目标分支迁移终点的错配中获得的最大影响。通过将MATE集成到磁诊断级联中,与之前的电路设计相比,我们在缩短12倍的分析时间内证明了相似的灵敏度。通过提供简单、快速、等温和非酶分析工作流程,我们的工作在使基于mnp的诊断更接近临床护理点设置方面取得了重大飞跃。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
9.10
自引率
0.00%
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0
审稿时长
10 weeks
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