In Situ Hand-in-Hand DNA Tile Assembly and CRISPR/Cas12a-based Ultrasensitive Detection for Leukemia Diagnosis,Sensors and Actuators B: Chemical

CRISPR/Cas technology has attracted increasing attention for use in accurate and sensitive nucleic acid detection, and has innovated the field of molecular diagnostics. Because it is programmability and shows high fidelity and effective signal amplification capabilities. Specifically, Cas12a has been evaluated through in-depth research because it triggers collateral trans-cleavage activity for the target. Here, we report a novel CRISPR/Cas12a fluorescent biosensor by in situ hand-in-hand DNA tile assembly for leukemia diagnosis, and it allows an active isothermal amplification for multi-channel collection of CRISPR/cas12a transducers for achieving three-stage signal amplification. When the target gene concentration was in a range from 1 fM to 50 nM, the fluorescence intensity exhibited an excellent linear relationship with the logarithm of the target gene concentration, and the detection limit reached 1 fM. Furthermore, good detection performance can still be maintained in actual sample analysis. The CRISPR/Cas biosensor for leukemia diagnosis is more advanced than the traditional CRISPR/Cas detection systems, which can be used in cancer gene analyses, kinship judgment, environmental monitoring, and so on.

中文翻译:








用于白血病诊断的原位手拉手 DNA 瓦片组装和基于 CRISPR/Cas12a 的超灵敏检测


CRISPR/Cas技术在准确灵敏的核酸检测中越来越受到关注,并在分子诊断领域进行了创新。因为它具有可编程性并显示出高保真度和有效的信号放大能力。具体而言,Cas12a 已通过深入研究进行评估,因为它会触发目标的旁系反式切割活动。在这里,我们报告了一种新型 CRISPR/Cas12a 荧光生物传感器,通过原位手拉手 DNA 瓦片组装用于白血病诊断,它允许主动等温放大,用于多通道收集 CRISPR/cas12a 传感器,以实现三级信号放大. 当目标基因浓度在 1 fM 到 50 nM,荧光强度与靶基因浓度的对数呈极好的线性关系,检测限达到1 fM。此外,在实际样品分析中仍能保持良好的检测性能。用于白血病诊断的CRISPR/Cas生物传感器比传统的CRISPR/Cas检测系统更先进,可用于癌症基因分析、亲属关系判断、环境监测等。








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