Label-Free Resonance Rayleigh Scattering Amplification for Lipopolysaccharide Detection and Logical Circuit by CRISPR/Cas12a-Driven Guanine Nanowire Assisted Non-Cross-Linking Hybridization Chain Reaction,Analytical Chemistry

Although the CRISPR/Cas system has pioneered a new generation of analytical techniques, there remain many challenges in developing a label-free, accurate, and reliable CRISPR/Cas-based assay for reporting the levels of low abundance biomolecules in complex biological samples. Here, we reported a novel CRISPR-derived resonance Rayleigh scattering (RRS) amplification strategy and logical circuit based on a guanine nanowire (G-wire) assisted non-cross-linking hybridization chain reaction (GWancHCR) for label-free detection of lipopolysaccharide (LPS). In the presence of a target, the protospacer-adjacent motif-inserted aptamer is rationally designed to specifically combine with LPS rather than Cas12a, suppressing the trans-cleavage activity of CRISPR/Cas12a and retaining the reporter probes to trigger non-cross-linking aggregation. Owing to the automatic hybridization chain reaction (HCR), in the presence of Mg2+, the released G-quadruplex sequence aggregated to assemble the G-wire superstructure through non-cross-linking. As a result, a dramatically amplified RRS intensity is observed, allowing for reporting LPS levels in a low detection limit of 0.17 pg/mL and a wide linear range among 1.0–100.0 ng/mL. Moreover, this reaction event is capable of programming to perform classical Boolean logic tree analysis, including basic logic computing and complex integrated logic circuits. This study comprehensively analyzed with respect to information flow, matter (molecular events), and energy (RRS), revealing the potential promise in designing of molecular-level “Internet of Things”, intelligent computing, and sensing systems.

中文翻译:








CRISPR/Cas12a 驱动的鸟嘌呤纳米线辅助非交联杂交链式反应用于脂多糖检测和逻辑电路的无标记共振瑞利散射放大



尽管 CRISPR/Cas 系统开创了新一代分析技术,但在开发一种基于 CRISPR/Cas 的无标记、准确和可靠的分析方法来报告复杂生物样品中低丰度生物分子的水平方面仍然存在许多挑战。在这里,我们报道了一种基于鸟嘌呤纳米线 (G-wire) 辅助非交联杂交链式反应 (GWancHCR) 的新型 CRISPR 衍生共振瑞利散射 (RRS) 放大策略和逻辑电路,用于脂多糖的无标记检测。脂多糖)。在存在靶标的情况下,原间隔区相邻基序插入的适配体被合理设计为与 LPS 而不是 Cas12a 特异性结合,抑制 CRISPR/Cas12a 的反式切割活性并保留报告探针以触发非交联聚合.2+,释放的G-四链体序列通过非交联聚合组装G-wire上层结构。结果,观察到显着放大的 RRS 强度,允许在 0.17 pg/mL 的低检测限和 1.0–100.0 ng/mL 的宽线性范围内报告 LPS 水平。此外,该反应事件能够编程执行经典的布尔逻辑树分析,包括基本逻辑计算和复杂的集成逻辑电路。本研究对信息流、物质(分子事件)和能量(RRS)进行了全面分析,揭示了分子级“物联网”、智能计算和传感系统设计的潜在前景。






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