Apart from gene editing capacity, the newly discovered CRISPR/Cas systems offer an exciting option for biosensing field because of their excellent target recognition accuracy. However, the currently constructed sensors are not only limited to nucleic acid analysis but also suffer from poor adaptability in complex samples and unsatisfying sensitivity. We herein introduce some advanced concepts to break through these bottlenecks. First, the sensing targets are extended by skillfully designing a functional DNA such as aptamer (for protein) and DNAzyme (for metal ion) to regulate the transduction of non-nucleic acid species and further activate the trans cleavage of CRISPR/Cas12a. Second, a boosting upconversion luminescent resonance energy is triggered by using a peculiar energy-confining notion, whereby the luminescence domain is intensively restricted in a very narrow space (~2.44 nm) and up to 92.9% of the green emission can be quenched by the approaching BHQ-1 modified reporters. Third, a bio-inspired periodic arrangement biomimetic chip (photonic crystal) is employed to selectively reflect the upconversion luminescence to achieve noteworthy signal enhancement (~35-fold). By utilizing very simple detection devices (a 980 nm portable laser and a smartphone), the CRISPR/Cas12a biosensor shows commendable sensitivity and specificity toward model targets (ATP and Na⁺, limits of detection are ~ 18 nM and ~0.37 μM, respectively). More importantly, the analysis of real complex samples demonstrate that the as-proposed platform can work as a powerful toolbox for monitoring the ATP fluctuation in single cell and point-of-care testing Na⁺ in human plasma, enabling a broad application prospect.

除了基因编辑功能外，新发现的CRISPR / Cas系统还具有出色的靶标识别精度，为生物传感领域提供了令人兴奋的选择。但是，当前构造的传感器不仅限于核酸分析，而且在复杂样品中的适应性差并且灵敏度不令人满意。我们在此介绍一些高级概念来突破这些瓶颈。首先，通过熟练地设计功能性DNA（例如适体（用于蛋白质）和DNAzyme（用于金属离子））来扩展感测靶标，以调节非核酸物质的转导并进一步激活反式CRISPR / Cas12a的切割。其次，通过使用特殊的能量限制概念来触发升频转换发光共振能量，从而将发光域严格限制在非常狭窄的空间（约2.44 nm）中，并且高达92.9％的绿色发射可以被淬灭。接近BHQ-1改装的记者。第三，采用具有生物启发性的周期性排列仿生芯片（光子晶体）来选择性地反映上转换发光，以实现显着的信号增强（约35倍）。通过使用非常简单的检测设备（980 nm便携式激光和智能手机），CRISPR / Cas12a生物传感器对模型目标（ATP和Na ⁺，检测限分别为〜18 nM和〜0.37μM。更重要的是，对实际复杂样品的分析表明，所提出的平台可以用作监测单细胞中ATP波动和人血浆中Na ⁺的即时检验的强大工具箱，从而具有广阔的应用前景。