CRISPR-Cas12a coupled with cyclic reverse transcription for amplified detection of miRNA
Xi Long, Jiacheng Li, Tong Luo, Hui Liu, Zhiwei Deng, Jiacheng Ding, Zan Gong, Yanjing Yang, Shian Zhong
Abstract
CRT. Afterward, the hairpin RT products could serve as activators to initiate the collateral cleavage activity of CRISPR-Cas12a. Due to the above two-stage amplification, this assay could detect miRNA at sub-femtomolar level (LOD, 0.201 fM). Since the sequence of target miRNA is double checked: first in the CRT and then in the CRISPR system, the proposed assay also shows an excellent specificity in detecting miR-21. Finally, with the usage of this assay, the sensitive assessment of miR-21 levels in human serum samples has been achieved and the disease human serum has been detected. Conclusively, CRISPR-CRT holds a great application prospective in the field of clinical molecular diagnosis.