Figure 7.

Validation of CRISPRD for detecting HPV DNA in human cell lines. (a) CaSki cells with an integrated HPV16 genome and HeLa cells with an integrated HPV18 genome were used to validate the triplex CRISPRD reaction. Human genomic DNA was extracted by a standard protocol of lysis, washing, and elution. (b) Validation of the CRISPRD reaction was performed using these cell lines. One master mix containing all reagents for a triplex reaction was prepared without adding the targets. DNA from each cell line was mixed to detect HPV16 and HPV18 targets. The reaction proceeded for 2 h (n = 3). HEK-293T cells (which do not contain HPV16 or HPV18 DNA) were used as negative control. The mean of the background-subtracted fluorescence is visualized in a heatmap. A total of 10 ng of total DNA extracted from the corresponding cell line was added to each reaction.