鉴于新兴的CRISPR/Cas12a检测技术具有目标选择性结合和切割活性，可以广泛应用于集成感知和执行系统。在此，我们详细介绍了一种通用方法，通过基于靶诱导球形核酸酶（SNA酶）网络的放大策略，快速激活CRISPR/Cas12a用于低丰度生物标志物检测。具体而言，设计了多功能的Y型探针和发夹探针（包含CRISPR/Cas12a激活体序列和DNA酶底物链的特定序列）构建SNA酶。目标识别导致Y型探针解离，释放DNA酶链，激活DNA酶，并伴随SNA酶自组装成SNA酶网络。有趣的是，与随机分散的SNA酶相比，SNA酶网络的反应动力学在α-甲基酰辅酶A环化酶（AMACR，前列腺癌生物标志物）的响应中提高了1.6倍，这归功于SNA酶网络对DNA酶的促进催化效率。借助这些特性，基于电化学发光（ECL）平台构建的生物传感器，通过将金银纳米簇（AuAgNCs）加载到金属有机框架-5（MOF-5）中，对AMACR显示出令人满意的检测性能，具有广泛的线性范围（0.001 μg/mL至100 μg/mL）和低检测限（1.0 × 10-4 μg/mL），在临床诊断中具有重要潜力。 版权所有 © 2023 Elsevier B.V.。保留所有权利。

Given the targeted binding ability and cleavage activity of the emerging CRISPR/Cas12a assay which transduces the target into its cleavage activity exhibited broadly prospective applications in integrated sensing and actuating system. Here, we elaborated a universal approach to quickly activate CRISPR/Cas12a for low-abundance biomarker detection based on the amplification strategy of a target-induced spherical nucleic acid enzyme (SNAzyme) network that could accelerate the output of activators. Specifically, multifunctional Y-shaped probes and hairpin probes (HPs, which contained the specific sequence of the activators of CRISPR/Cas12a and the substrate chain of DNAzyme) were rationally designed to construct SNAzyme. Target recognition induced disassembly of the Y-shaped probes, which released DNAzyme strands to active DNAzyme and accompanied by SNAzyme self-assembly into SNAzyme network. Interestingly, compared with randomly dispersed SNAzyme, the reaction kinetics of the SNAzyme network enhanced 1.6 times in response to Α-methyl acyl-CoA racemase (AMACR, a biomarker for prostate cancer), which was attributed to the promoted catalytic efficiency of DNAzyme by the confined SNAzyme network. Benefiting from these, the prepared biosensor based on electrochemiluminescence (ECL) platform by loading AuAg nanoclusters (AuAgNCs) into metal-organic framework-5 (MOF-5) exhibited satisfying detection performance for AMACR with a wide linear range (0.001 μg/mL to 100 μg/mL) and a low detection limit (1.0 × 10-4 μg/mL, which exhibited significant potential in clinical diagnoses.Copyright © 2023 Elsevier B.V. All rights reserved.