用于无分裂自催化扩增的可编程类PCR非酶DNA分子电路

Programmable PCR-like Nonenzymatic DNA Molecular Circuit for Split-Free Autocatalytic Amplification.

作者信息

Li Ting, Lau Tat San, Li Junyou, Hu Kaiqi, Lee Man Lung, Chen Pin You, Wang Chi Chiu, Li Hung-Wing

机构信息

Department of Chemistry, The Chinese University of Hong Kong, Shatin New Territories 999077, Hong Kong SAR, China.

Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Shatin New Territories 999077, Hong Kong SAR, China.

出版信息

Anal Chem. 2025 Jun 10;97(22):11847-11855. doi: 10.1021/acs.analchem.5c01612. Epub 2025 May 29.

Abstract

Nonenzymatic autocatalytic DNA circuits, capable of exponential signal amplification, exhibit superior amplification efficiency compared to traditional single or cascade DNA amplification circuits, which offer only linear or quadratic signal amplification. However, autocatalytic DNA circuits are currently limited by complicated splitting designs, low autocatalytic efficiency, the need for additional probes, and a lack of universal design principles. Herein, we developed a PCR-like split-free autocatalytic amplification (SAA) DNA circuit with a simple design but extraordinary autocatalytic efficiency for effective biosensing. The SAA system could consecutively perform multiple cycles of a three-step process upon initiation, target recognition, replicating, and recycling, which is programmed to mimic the basic three reaction steps of PCR and constantly yield numerous new split-free target replicates to expedite the whole reaction, ultimately producing an exponentially amplified signal. The PCR-like sigmoidal kinetics, the ability to accurately execute the programmed instructions, and the high autocatalytic capability of the SAA system are demonstrated, which achieves the same target replication as the PCR but without the need for enzymes and precise control of temperature. The SAA circuit, characterized by exponential signal amplification, simple design, and minimal components, offers a promising approach for developing highly efficient universal DNA circuits. This enables the analysis of low-abundance biomarkers with minimal signal leakage, holding significant potential for biochemical research and clinical diagnosis.

摘要

非酶促自催化DNA电路能够进行指数信号放大,与传统的单链或级联DNA放大电路相比,具有更高的放大效率,后者仅提供线性或二次信号放大。然而,自催化DNA电路目前受到复杂的拆分设计、低自催化效率、对额外探针的需求以及缺乏通用设计原则的限制。在此,我们开发了一种类似PCR的无拆分自催化扩增(SAA)DNA电路,其设计简单但自催化效率非凡,可用于有效的生物传感。SAA系统在启动、靶标识别、复制和循环时可连续执行三步过程的多个循环,该过程被设计为模拟PCR的基本三个反应步骤,并不断产生大量新的无拆分靶标复制品以加速整个反应,最终产生指数放大信号。证明了SAA系统具有类似PCR的S形动力学、准确执行编程指令的能力以及高自催化能力,其实现了与PCR相同的靶标复制,但无需酶和精确的温度控制。SAA电路具有指数信号放大、设计简单和组件最少的特点,为开发高效通用DNA电路提供了一种有前景的方法。这使得能够以最小的信号泄漏分析低丰度生物标志物,在生化研究和临床诊断方面具有巨大潜力。

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