Figure 4 | Scientific Reports

Figure 4

From: Single crossover-mediated targeted nucleotide substitution and knock-in strategies with CRISPR/Cas9 system in the rice blast fungus

Figure 4

Single crossover-mediated reporter gene knock-in at the SDH locus with CRISPR/Cas9. (a) Schematic representation of the single crossover-mediated GFP knock-in at the SDH locus. The start and stop codons were deleted from SDH gene and the silent mutations were introduced at the CRISPR/Cas9 target site of this gene. GFP was fused to mutated SDH C-terminus. (b,c) GFP fluorescence in the wild type (b) and GFP-tagged transformant. (c) BF: Bright-field image, GFP: epifluoresence image. The bars are 500 µm. (d) The efficiencies of GFP knocked-in transformant. Total colony: number of hygromycin B-resistant colonies obtained from repeated experiments. Fluorescent colony: number of hygromycin B-resistant colonies presenting the GFP fluorescence. Efficiency: percentage of total GFP fluorescent colonies. (e) Sequences of the CRISPR/Cas9 target region in the wild type, donor vector, and fluorescent colonies of the transformants.

Back to article page