Department of Biochemistry and Molecular Biology, Shanxi Medical University, 56 Xinjian South Road, Taiyuan, 030000, Shanxi, P.R. China.
College of Safety and Emergency Management and Engineering, Taiyuan University of Technology, Taiyuan, 030000, Shanxi, China.
Mikrochim Acta. 2024 Jun 14;191(7):395. doi: 10.1007/s00604-024-06476-0.
With their regulated Boolean logic operations in vitro and in vivo, DNA logic circuits have shown great promise for target recognition and disease diagnosis. However, significant obstacles must be overcome to improve their operational efficiency and broaden their range of applications. In this study, we propose an Exo III-powered closed-loop DNA circuit (ECDC) architecture that integrates four highly efficient AND logic gates. The ECDC utilizes Exo III as the sole enzyme-activated actuator, simplifying the circuit design and ensuring optimal performance. Moreover, the use of Exo III enables a self-feedback (autocatalytic) mechanism in the dynamic switching between AND logic gates within this circulating logic circuit. After validating the signal flow and examining the impact of each AND logic gate on the regulation of the circuit, we demonstrate the intelligent determination of miR-21 using the carefully designed ECDC architecture in vitro. The proposed ECDC exhibits a linear detection range for miR-21 from 0 to 300 nM, with a limit of detection (LOD) of approximately 0.01 nM, surpassing most reported methods. It also shows excellent selectivity for miR-21 detection and holds potential for identifying and imaging live cancer cells. This study presents a practical and efficient strategy for monitoring various nucleic acid-based biomarkers in vitro and in vivo through specific sequence modifications, offering significant potential for early cancer diagnosis, bioanalysis, and prognostic clinical applications.
基于其在体外和体内的调控布尔逻辑运算，DNA 逻辑电路在目标识别和疾病诊断方面显示出巨大的应用潜力。然而，为了提高其操作效率并拓宽其应用范围，仍需克服重大障碍。在本研究中，我们提出了一种基于 Exo III 的闭环 DNA 电路（ECDC）架构，该架构集成了四个高效的 AND 逻辑门。ECDC 仅使用 Exo III 作为唯一的酶激活执行器，简化了电路设计并确保了最佳性能。此外，Exo III 的使用使得在这个循环逻辑电路中，AND 逻辑门之间的动态切换能够实现自我反馈（自催化）机制。在验证了信号流并研究了每个 AND 逻辑门对电路调节的影响之后，我们在体外使用精心设计的 ECDC 架构展示了对 miR-21 的智能检测。所提出的 ECDC 对 miR-21 的线性检测范围为 0 到 300 nM，检测限（LOD）约为 0.01 nM，超过了大多数报道的方法。它还对 miR-21 的检测表现出优异的选择性，并具有识别和成像活癌细胞的潜力。本研究通过特定序列修饰，为体外和体内监测各种基于核酸的生物标志物提供了一种实用高效的策略，有望用于早期癌症诊断、生物分析和预后临床应用。
