CRISPR-Cas12a enhanced rolling circle amplification method for ultrasensitive miRNA detection

By Gong Zhang, Lei Zhang, Jing Tong, Xianxian Zhao, Jianlin Ren. 2020

123 citations recorded by the source corpus.

Abstract MicroRNAs (miRNAs) detection with high specificity and sensitivity received abundant attention because miRNAs have been reported to play a vital role in pathological development of many diseases and regarded as potential biomarkers for the diagnosis and prognosis of diseases. We reported here a highly specific method for molecular exosomal miRNAs detection in constant temperature by integrating the advantages of CRISPR/Cas system and rolling circular amplification (RCA) techniques. Especially, the proposed strategy was demonstrated to obtain a high sensitivity attributed to the dual-specific recognition from miRNA-padlock initiated RCA and CRISPR-Cas12a-triggered specific cleavage. Eventually, the proposed strategy showed a sensitivity of 34.7 fM which was robust enough for exosomal miRNA detection and obtained a high consistency with reverse transcription quantitative polymerase chain reaction (RT-qPCR), revealing the potential of developing a universal molecular detection platform for the screening, diagnosing, and prognosis prediction of multiple diseases.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/fb853c5fda1d2f922c96e59177074c62d4577b9a