级联 CRISPR/cas 实现了无需扩增的 microRNA 感应，具有 fM 级灵敏度和单碱基特异性。
Cascade CRISPR/cas enables amplification-free microRNA sensing with fM-sensitivity and single-base-specificity.
机构信息
MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, P. R. China.
出版信息
Chem Commun (Camb). 2021 Jan 7;57(2):247-250. doi: 10.1039/d0cc06412b. Epub 2020 Dec 11.
Existing CRISPR/cas-based biosensors usually improve sensitivity by target amplification, which is time-consuming and susceptible to impurities in complex biofluid. Herein, this is the first time a cascade CRISPR/cas (casCRISPR) system has been developed, which can provide a detection limit of 1.33 fM (∼1000 times lower than direct Cas13a-based miRNA detection) and single-base resolution for miR-17 detection without resorting to target amplification. casCRISPR can also be applied to detect miRNA in complicated cell extracts and serum samples. Overall, casCRISPR will provide a heuristic idea for CRISPR/cas based biosensing, and could be a promising tool for miRNA diagnostics.
现有的基于 CRISPR/cas 的生物传感器通常通过目标扩增来提高灵敏度，但这种方法既耗时又容易受到复杂生物流体中杂质的影响。在此，首次开发了级联 CRISPR/cas（casCRISPR）系统，可在不进行目标扩增的情况下，实现对 miR-17 的检测，其检测限低至 1.33 fM（比直接基于 Cas13a 的 miRNA 检测低约 1000 倍），且具有单碱基分辨率。casCRISPR 还可用于检测复杂细胞提取物和血清样本中的 miRNA。总的来说，casCRISPR 为基于 CRISPR/cas 的生物传感提供了一个启发式的思路，有望成为 miRNA 诊断的一种有前途的工具。
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