Integrating CRISPR-Cas12a with a DNA circuit as a generic sensing platform for amplified detection of microRNA
Shuang Peng, Zhen Tan, Siyu Chen, Chunyang Lei, Zhou Nie
Abstract
-cleavage activity of CRISPR-Cas12a for further signal amplification. Such rational integration provides a generic assay for the effectively amplified detection of miRNA biomarkers. By simply tuning the variable regions in the CHA modules, this assay achieves sub-femtomolar sensitivity for different miRNA biomarkers, which improves the detection limit of CRISPR-Dx in the analysis of miRNA by 3-4 orders of magnitude. With the usage of the proposed assay, the sensitive assessment of miR-21 levels in different cancer cell lines and clinical serum samples has been achieved, providing a generic method for the sensitive detection of miRNA biomarkers in molecular diagnosis.
Topics & Concepts
CRISPRmicroRNAComputational biologyDNAComputer scienceBiologyGeneticsGeneCRISPR and Genetic EngineeringAdvanced biosensing and bioanalysis techniquesRNA Interference and Gene Delivery