Fig. 2: Construction of a Crispr-driven AIEgen lighting-up readout. | Nature Communications

Fig. 2: Construction of a Crispr-driven AIEgen lighting-up readout.

From: CRISPR/Cas-mediated “one to more” lighting-up nucleic acid detection using aggregation-induced emission luminogens

Fig. 2

a Excitation spectra (dotted line) and emission spectra (solid line) of FAM (green) and TPBT (red). b Photobleaching resistance of FAM and TPBT. Normalized fluorescence intensity of FAM and TPBT at various time points, during the 60 min successive scanning of the respective excitation beam. c, d Proof of concept of S1 nuclease-driven AIEgen lighting-up readout. c Fluorescence emission spectrum of Q-ssDNA-FAM, free TPBT, TPBT mixed with ssDNA or Q-ssDNA, and released FAM or TPBT after splitting ssDNA by S1 Nuclease. d Fluorescence emission spectrum of free TPBT, TPBT mixed with dsDNA, TPBT mixed with Q-dsDNA or Q-dsDNA-Q and released dsDNA/TPBT after splitting ssDNA linker by S1 Nuclease. The synthetic norovirus ORF2 gene DNA sequence was used as the model analyte to construct the Crispr/Cas12a sensing system. Schematic illustration of CrisprAIE based on ssDNA/AIEgen (e) Q-ssDNA/AIEgen (f), Q-dsDNA/AIEgen (g) and Q-dsDNA/AIEgen-Q (h) reporters. Relative fluorescence intensity obtained from amplification-free CrisprAIE assay with different concentrations of synthetic DNA targets based on ssDNA/AIEgen (i), Q-ssDNA/AIEgen (j), Q-dsDNA/AIEgen (k), and Q-dsDNA/AIEgen-Q (l) reporters. Data are mean ± s.d.; n = 3 repeated tests.

Back to article page