Fig. 1

Comparison of the expression level and the activity of the CRISPR-Cas12a systems using different promoters. a Schematic of the CRISPR-Cas12a systems driven by different promoters (CAG, EF1a Core, CMV, and PGK). b Western blot showing the expression levels of the Cas-protein nucleases (anti-HA) driven by different promoters. Blank, MCF7 without transfection. c FACS detected the mCherry expression level driven by different promoters in MCF7 cells. The left panel showing the transfection efficiency with the input of the same DNA while the right panel displaying the expression levels by calculating the mean of the fluorescence (mCherry) intensity. Mean values are presented with SEM, n = 3 independent experiments. d FACS analyses of the editing activities of the CRISPR-Cas12a systems with different promoters with four crRNAs that targeted the mNeonGreen in the HEK293T KI mNeonGreen reported cell line. The editing efficiency was determined as the proportion of mNeonGreen negative cells within the Cas-nucleases transfected cells (mCherry-positive). crRNA1/2/3/4, crRNAs targeting mNeonGreen. Mean values are presented with SEM, n = 3 independent experiments. Blank, cells without transfection. NC, cells transfected with a non-targeted crRNA