sfGFP gene knockout in HEK293T cells transfected with the LAC12aGE machinery and the fluorescent dual reporter plasmid. Panel I: Confocal microscopy images corresponding to the fluorescence of sfGFP and mCherry in HEK293T cells. Entry i, Negative control (Neg.) transfected with Cas12a plasmid in the absence of crRNAsfGFP. Entry ii, Positive control (Pos.) transfected with Cas12a plasmid and wt-crRNAsfGFP. Entry iii, Cells transfected with Cas12a plasmid/OFF-crRNAsfGFP in the absence of UV light (UV). Entry iv, Cells transfected with Cas12a plasmid/OFF-crRNAsfGFP in the presence of UV light (+UV). Scale bar, 50 m. Panel II: The relative ratios of sfGFP to mCherry derived from the integrated fluorescence intensities of sfGFP and mCherry displayed in panel I. Panel III: Disruption efficiencies of sfGFP by the Cas12a positive control and the LAC12aGE machinery, where the sfGFP disruption efficiency (%) by the Cas12a is defined as (FluNeg.–FluPos.)/FluNeg. 100%, and the disruption efficiency (%) by the LAC12aGE is defined as (FluUV Flu+UV)/FluUV 100%.
