Fig. 3

Validation of the CRISPR/Cas12a system. A The secondary structure sequences of the six crRNAs of the LbCas12a and FnCas12a protein; (B) crRNAs obtained after in vitro transcription purification, lane M: DL2000 DNA marker; lane 1: LbcrRNA1, lane 2:LbcrRNA3, lane 3: FncrRNA1, lane 4: FncrRNA3; (C) Fluorescence kinetics compared the sensitivity of six individual crRNAs to detect circRNA, and the concentration of circMUC16 was set to 50 pM; (D) Heatmap of end-point fluorescence values of circRNA from six individual crRNAs; (E) Verify the feasibility of CRISPR system to detect circRNA, fluorescence kinetics of FnCas12a/FncrRNA1, LbCas12a/LbcrRNA1 and FnCas12a/FncrRNA1 + LbCas12a/LbcrRNA1 in the presence and absence of target cDNA; (F) The heatmap shows the end point fluorescence value to validate the feasibility analysis of CRISPR detection system