Cascade CRISPR/cas enables amplification-free microRNA sensing with fM-sensitivity and single-base-specificity

作者：Yong Sha, Ru Huang, Mengqi Huang, Huahua Yue, Yuanyue Shan, Jiaming Hu, Da Xing · 发表于：Chemical Communications · 年份：2020 · DOI：10.1039/d0cc06412b · 被引用次数：135 · 研究领域：CRISPR and Genetic Engineering、Advanced biosensing and bioanalysis techniques、RNA Interference and Gene Delivery

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.