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Applied Biological Chemistry
Fig. 2 | Applied Biological Chemistry

Fig. 2

From: Optimized protocols for protoplast isolation, transfection, and regeneration in the Solanum genus for the CRISPR/Cas-mediated transgene-free genome editing

Fig. 2

PEG-based protoplast transfection for gene edition. A An overview of the PEG-based protoplast transfection procedure. Transfection can be confirmed by visualizing a marker protein, GFP. B The viability of tomato protoplasts was observed under a fluorescence microscope. Protoplasts with green fluorescence were viable. Scale bars indicate 50 μm. Protoplasts exhibiting a perfect round shape were counted as viable protoplasts (n = 6; *p < 0.05). C Comparison of PEG-mediated protoplast transfection and Agrobacterium-mediated transfection targeting the ALS2 gene in tomato cells using the CRISPR/Cas9 system. The target sequences are in bold, and the edited regions are in red. The black underline indicates the gRNA sequences, and the red letters indicate the PAM sequence

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