Fig. 1

Schematic illustration of protoplast isolation and CRISPR/Cas9-mediated genome-editing steps via Lipofectamine. Protoplasts isolated from in vitro grown cell suspension (A). Protoplast layer represents protoplasts between solutions of sucrose 25% and mannitol 13%. Bright field of protoplast 1 h after isolation. Mixing of lipofectamine and plasmid constructs targeting NPR3 and protoplast transfection (B). Characterization steps by monitoring EGFP expression and cas9 presence in the genomic DNA, T7EI assay and sanger sequencing and gene expression analysis (C). GFP-expressed cell was monitored by a confocal laser scanning microscope. The figure was created in BioRender.com