Analysis of the Effectiveness of Crispr-Editing of the GEX2 Gene by Ribonucleoprotein Complexses in Maize Protoplasts

The GEX2 protein is expressed in the maize gamete membranes and necessary for gamete membranes contact (adhesion). Knockout of GEX2 gene, presumably, can lead to impaired fertilization and, as a result, to the haploid embryo formation. The aim of the study is to analyze the efficiency of CRISPR/Cas9 editing of the GEX2 gene after PEG-mediated transfection of maize protoplasts by ribonucleoprotein (RNP) complexes with different sgRNA. For the first time, the RNP complexes with different sgRNA to the GEX2 gene have been created. The effectiveness of CRISPR/Cas9 editing of the GEX2 gene have been proven on protoplasts and reaches 10.7%, depending on the sgRNA, level and thesgRNA:Cas9 ratio in the RNP complex.