Fig. 2.

Optimization of the backbone plasmid amount used in LION and validation of LION CRISPR vector for gene editing by C-Check. a Diagram depicting the working principle of our C-check system. SSA single-strand annealing, NHEJ non-homologous end joining. The AsRed expression cassette is used as a background fluorescence for normalization of the transfection efficiency. b Left: an example of the C-Check assay in cells transfected with or without CRISPR/Cas9 plasmids. Right: FCM patterns of the C-Check efficiency test for different amounts of backbone used in LION. mock denotes cell samples transfected with the same total amount of pUC19 plasmid. c FCM quantification of C-Check cleavage efficiency between LION and pure CRISPR vectors (n = 3). w/o without. Asterisk (**) indicates a p value less than 0.01