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可编程的分裂DNA酶调节器通过Allosteric合作激活mRNA电化学发光生物感知

Liu-Qing Tan¹, Wei-Jia Zeng¹, Qiao-Lin Chen¹

¹MOE Key Laboratory of Luminescence Analysis and Molecular Sensing, College of Chemistry and Chemical Engineering, Institute of Developmental Biology and Regenerative Medicine, Southwest University, Chongqing 400715, P. R. China.

Analytical chemistry

|March 3, 2025

概括

一种新型的全合作激活策略为DNA酶调节器 (STATER) 编程,用于敏感检测IL-6 (IL-6) mRNA. 这种方法提高了准确性,并最大限度地减少了复杂的生物样本中的假阳性.

科学领域:

生物技术和生物传感技术
分子生物学分子生物学
纳米技术纳米技术

背景情况:

DNA酶为信号传导提供了可编程性和稳定性,但在催化效率和意外激活方面面临挑战.
现有的DNA酶调节器通常依赖于单个效应器激活,限制了复杂生物矩阵中的特异性和灵敏性.
准确检测诸如白蛋白-6 (IL-6) mRNA之类的生物标志物对于理解生物过程和疾病状态至关重要.

研究的目的:

开发一种新的DNA酶调节器,STATER,采用全合作激活策略来增强生物感知.
通过克服传统DNA酶系统的局限性,实现IL-6mRNA的敏感和准确的电化学发光 (ECL) 检测.
确定IL-6mRNA检测的激活值,减少复杂生物样本中的非特异性激活.

主要方法:

设计了一个使用DNA四面体支架的STATER,配有T形发针探头 (TP) 和辅助发针探头 (HP).
纳入APurinic/apyrimidinic内核酶1 (APE1) 识别位点和IL-6 mRNA结合区域的TP.
在黄金纳米集群平台上实施了一种全合作激活机制,涉及部分酶组装和ECL信号生成的托克交换位移.

主要成果:

通过ECL生物传感器,STATER证明了对IL-6mRNA的敏感和选择性检测.
生物传感器显示IL-6mRNA的线性检测范围从1×10-13到1×10-7M.
达到3.26×10−14M的低检测极限,表明高灵敏度和特异性.

结论:

全合作激活策略有效地提高了DNA酶调节器性能,用于特定的分析物检测.
开发的基于STATER的ECL生物传感器显示出在复杂的生物系统中对敏感和准确的IL-6mRNA量化具有重大潜力.
这种方法为开发各种生物分析物的先进生物传感器提供了一个强大的平台.

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